Joining 3.5 Billion Years of Microbial Invention featuring biologist J. Craig Venter.
Biologist, author and businessman Craig Venter discusses his work mapping and synthesizing genomes. Venter recalls his work mapping the human genome and expands on his current work which includes categorizing new genes and species of microbes from ocean water. Venter also explains how microbial research can be used for metabolic engineering and alternative energy sources.
Stewart Brand is co-founder and president of The Long Now Foundation and co-founder of Global Business Network. He created and edited the Whole Earth Catalog (National Book Award), and co-founded the Hackers Conference and The WELL. His books include The Clock of the Long Now; How Buildings Learn; and The Media Lab. His most recent book, titled Whole Earth Discipline, is published by Viking in the US and Atlantic in the UK.
J. Craig Venter
J. Craig Venter, Ph.D., is regarded as one of the leading scientists of the 21st century for his numerous invaluable contributions to genomic research. He is Founder, Chairman, and CEO of the J. Craig Venter Institute (JCVI), a not-for-profit, research organization with approximately 300 scientists and staff dedicated to human, microbial, plant, synthetic and environmental genomic research, and the exploration of social and ethical issues in genomics.
Dr. Venter is also Founder and CEO of Synthetic Genomics Inc (SGI), a privately held company dedicated to commercializing genomic-driven solutions to address global needs such as new sources of energy, new food and nutritional products, and next generation vaccines.
Scientific study of microorganisms, a diverse group of simple life-forms including protozoans, algae, molds, bacteria, and viruses. Microbiology is concerned with the structure, function, and classification of these organisms and with ways of controlling and using their activities. Its foundations were established in the later 19th century, with the work of Louis Pasteur and Robert Koch. Since then, many disease-causing microorganisms have been identified and means of controlling their harmful effects have been developed. In addition, means of channeling the activities of various microorganisms to benefit medicine, industry, and agriculture have been discovered. Molds, for example, produce antibiotics, notably penicillin. See alsobacteriology, genetic engineering.
This stuff is genuinely on the frontiers of science. CO2 as a feedstock to create "fourth generation fuel" is esp interesting. I'm guessing it won't happen tomorrow or the next day but it does seem that this stuff is legitimately on the horizon.
whatevs, CO4E. The section on the number of microbes around us at any given time -- not to mention the number we absorb by sitting in a room with other people for an hour-- is mind blowing. (but there is a bit of an 'ewwww' factor.)
Good evening. (Applause) Thank you for coming I'm Stewart Brand from the Long Now Foundation Ishould say that most of you have these question cards and in this semi-darkness of the theater. ItÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢shelpful when youÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ve written out a question to come up to the front for Kevin Kelly and me to look attoo. Later address the speaker with itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s helpful if you wave it. So, that the guys in the yellow hatscan see it come collect it and bring it up. And you know write questions anytime during the talk,during the Q&A thatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s the interactive part.Now, the Long Now Foundation is called the long now because we have in mind the next 10,000 yearsin the last 10,000 years and Peter Schwartz here is the one who came up with that number because10,000 years ago there was a biotech revolution having to do with mainly plants and then animals thatwe eat and use. Now, we started genetic modifications, genetic engineering in a big way and humanitytook a swerve that itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s still getting used to.And in about 150 years, another biotech revolution happened with biomedicine and humans started tobe able to control their health, be able to control their birth rate and we get to the 21st century whichregarded by many as the century of biology, which to a large extent means the century ofbiotechnology. And so these revolutions that weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ve seen seem to be on a kind of biotech MooreÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s Lawof getting more and more frequent and happening now in one lifetime instead of it millennia at a time.In the current acceleration of genetic science and genetic engineering is as often the case being carriedby very sharp individuals. Scientist who find the right problems, find the right people to work with,find the right funding and swarm ahead. And really the exemplary case of that these days is CraigVenter. Please welcome him.Well thank you Stewart itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s certainly nice to be here with The Long Now Foundation taking a longview hopefully forward with Humankind. I think itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s very much an open question, but weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re trying tosee if we can change some of the equations maybe to help that process along. Again, talking about twophases of information tonight, the first phase is gathering information. Actually referred to it and inmy case is reading the genetic code and then weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ll turn to how now weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re using that information tochange evolution, to change biology, to change hopefully some parts of society with writing the geneticcode. And IÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ll talk about the early stuffs of that. All this is happening I a pretty short period of time.WeÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ve sequenced the first genome of a living species Haemophilus influenzae in 1995. Before themwe developed techniques for scaling up gene discovery. But weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ve only had complete genomes ofliving organisms for a relatively short period of time. ItÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s changing our view of looking at biology.We did two genomes in 1995 and IÃƒÂ¢Ã¢â€šÂ¬Ã‹Å“ll about the second one in the later phases of this had very differentsize genomes, very different characteristics. But after those first two that we had to fund ourselves westarted getting a lot of government funding to scale up the process. We got funding to do most humanpathogen. WeÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ve moved onto plants than simple animals. Then more complex ones such the fruit flies and people.The big change in technology after 1995 came surprisingly only four to five years later with a fruit flygenome. Haemophilus was 1.8 million letters of genetic code, Drosophila 180 million. And they wereboth done in the same four months time period. And 9 months later we had the three Billion first draftof the human genome. Computational aspects weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re very important. We actually had to build thethird largest computer in the world to do this calculation. Now itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s not hard to do with much morestandard computers. One and half tera-ops in 1999 was a huge computer. Today, we have things 10,20, 30 and 100 times of that size.Just hits last year we published the first complete human genome. This was the diploid genome, sort ofhad both sets of chromosomes from both parents. What was done in 2000 and 2001 ended only beinghalf the job. The public effort only got it half right because they set pout only to do half. Taking theother half would e easily discernible. The project of Solera actually tried to do the complete genomebut we the mistake of overreaching we tried to do the genome of five individuals. There were threemen and two women. No there was three women and two men. It sounds like a San Francisco movie,but because of the diversity, when we assembled the other genome. We ended up subtracting out allthe major variation between them show the message and many you will remember it.In 2000-2001 as we all have the same set of genes and we all differ from each other only in one out of athousand letters of genetic code. And this started all kinds of people down certain tracks. We have apretty large industry now just to measure those one of thousand differences but it turns out weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re all 1to 3 percent different from each other. So itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s a much greater variation than we imagined. To put thatin context, we thought we differed from chimpanzees only by 1.27%. So youÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re probably hoping thatnumber changed as well or we really have explaining to do about evolution. It turns out weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re 4-5%different from chimps. And so this individual variation was totally missed. The two sets ofchromosomes that each of you have. Each of those sets of chromosomes they donÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢t naturally have thesame gene that the other pair has. For example thereÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s a major gene associated with detoxifyingenvironmental toxins that of the Caucasian population a third have no copied of this gene. Anotherthird have open copy and another third have two copies. But even when you have two copies thereÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢smajor variation of these. So itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s hard to find two of us with the exact same genetic code and we have alot of complexity. I think itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s important that we know it now because it means we can move towardsgetting the right answers.In this first genome almost half the genes had a major variant in them and this certainly going toconfound any body who was hoping to get simple interpretation of the human genetic code. So wehave all kinds of companies promising that now for a thousand dollars. But itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s only a partial peek atwhat might be there. Seventy five percent of the letters the genetic code there variant between in anytwo of us. DidnÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢t show in the single nucleotide polymorphisms at that people were measuring.So, were scaling it out now to do 10,000 human genomes over the next decade. Hopefully, it won'ttake 10,000 years to it. But maybe only 10 and we will finally have the first chance to understandwhatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s genetic, whatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s nature and what's nurture. Not only with the genetic code but were trying tocollect intense extensive phenotypic information on all these individuals that were going to besequencing and thatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s a much greater challenge when you think about all the information that mightdescribe you or your lives. How you look, how you think, your body structure, you organ structure,your metabolism all those things differ dramatically with each of us and trying to describe that makessequencing the genome look like the easy part.We published our data in the public access of journal biology. You can download this 300 megabytefile, if you want to. ItÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s a zoomable version of the human genome that shows the types and extent ofhuman variation and hopefully in a short time youÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ll be able to get the map of your genome this way.Working with the X PRIZE Foundation, we now have a 10-million-dollar price for who ever gets thetechnology going to the extent that we can sequence an individual genome for a thousand dollars in avery short period of time. So, weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re hopeful that weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ll be giving out this 10 million dollars in the next5 years, if youÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re looking for a little spare change. You might try inventing something worthwhile.WeÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re releasing a new browser for the genome that allows it to be looked at down to the sequence filelevel as people for example get data from 23 in media or other places. They can look at actually acomplete genome to try and put it some kind of context and find out how much is missing from those early test.Now, when we finish sequencing the human genome many people were hoping I would retire, butinstead we looked around for what we thought were the most important projects in science to doparticularly with that the technology that we developed for reading the genetic code to try and tackle.And it was clear to us that trying to look at the environment not only the extensive environment aroundus that helped influence our genes. But the broader environment to see if we could use DNA sequenceas a new tool to see who we are sharing the planet with. ThereÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s been this statistic for a long time thateach milliliter of seawater has a million bacteria and 10 million viruses. But nobody really knew orwhat that meant and whether there was any diversity in that. And so we decided to do an experiment inthe Sargasso Sea just taking a barrel of seawater filtering out all the organisms in it. Isolating the DNAfrom them on mass and sequencing it and just from one barrel of water we stopped sequencing after wehad close to 1.4 million new genes. Maybe as 40,000 new species that had never been seen before. Sothe technique obviously worked because in fact people thought weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢d find little or no life in theSargasso Sea because it was supposed to be a desert with no nutrients in it.Again, IÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ll show you why and how these organisms survived in a minute, but we decided to lookfurther and we started the Sorcerer II expedition. I had always been looking for an excuse to sailaround the world on my own vessel. And so we decided we would follow great scientific expeditionssuch as the Challenger expedition and sail around the world and taking samples every 200 miles. Andsequencing everything we could find to see if we could generate a different view of life. And we wereabsolutely stunned with what we found and that was a special issue of PLoS that was published last year.This is the route that we followed, as with the Challenger expedition, we started in Halifax, went donethe Eastern seaboard into the Caribbean Sea then down between Mexico and Cuba to the PanamaCanal. Through the Panama Canal and down first Cocos Island and then to the Galapagos and thatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢sthe dot you see here in blue. ThatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s what was covered in the special issue of PLOS of biology and thatcovered over 6 million new genes more than doubling the number of genes known from all science upto that point. And weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ve now been analyzing the samples from the rest of the globeAfter Galapagos, we left and sailed to the Marquesas where we are promptly arrested because therewas a debate between the French government and the French Polynesia government as who had rightsover the microbes that are there. You may not understand this but this is a physical map if you look ata political map in the Caribbean Sea for example there's no international waters. Every drop of water isclaimed by one or more countries and you need permission from them to take a scientific example.Its okay to fish on most of those places you can shoot marine mammals you could do whatever youwant. But if youÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re asking scientific questions itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s considered extremely dangerous and you can getarrested for it. In fact, when the organisms in the ocean are out in the middle of the Pacific Ocean,theyÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re in international waters and they belong to nobody or to everybody. As soon as the one-knotcurrent that goes across the Pacific carries them into French Polynesia all those microbes instantlybecomes French genetic heritage. (Laughter) And they're willing to defend it to the extreme.We finally got out of there and took samples and spent some time in Australia then went across theIndian Ocean. We got halfway across and we and we stopped in Chagos Island, which is near whereour country has a B-52 Air Base and weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re arrested again by the British this time because we wouldtake samples and understand science in that water. So, science is something and organisms that aregreatly protected around the world. And to be able to publish this date in the public databases we hadto put geographic GPS coordinates on every DNA sequence.So, for example is somebody makes a discovery using the sequences that we found in Australia andyou want to commercialize those you have to contact the Australian government and negotiatesomething thatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s not clear what. So science is much more complicated today, but the experiments werevery simple. We just simply filtered seawater through different size filters. We could then just takethose filters put them in a freezer. When we got into a port, we put them in a FedEx package withsome dry ice and sent them back to the lab in Rockford, Maryland where all the DNA was isolated atonce from and sequenced and then reconstructed in the computer.And when we reconstructed it, we are amazed with what we found. For example, every 200 miles,85% of the sequences and the organisms are unique. The ocean is not homogenous mixture. Samplesthat weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ve taken off of San Francisco Bay will be very different than the ones off of Los Angeles andoff Seattle and Oregon. In fact if you look at the ÃƒÂ¢Ã¢â€šÂ¬Ã¢â‚¬Å“ so theyÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re red or dark here if you color blind iswarm water the blue light colors is cold water. And even the 3% of sequences that assemble acrossmore than one site. TheyÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re an absolute distinction between a warm in cold water. And some of thesechange on an annual basis with weather.Other things that affect it or sunlight, But ÃƒÂ¢Ã¢â€šÂ¬Ã¢â‚¬Å“ and nutrients we can tell simply from a sample of seawater.Looking in the DNA in that water, where that water came from in the world? And for example, withall the ships that had come and gone from San Francisco Bay. If a tanker comes in and offloads itcargo, it fills up its hold with seawater. And weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ll go to its next port, dump all that seawater and takeon new cargo. So, look at all the ships that come and go from San Francisco Bay each day and imaginewith a million bacteria and 10 million viruses per milliliter. How these environments are constantlybeing challenged and shifted.One of the biggest discoveries weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ve made early on was that these organisms we discovered in theocean had photoreceptors. Molecules very to our own visual pigments and almost everyone in the topparts of the ocean have these photoreceptors. The blue segment at the bottom of the slide is our priorknowledge of photoreceptors before this expedition. And these are very deep branching. A lot ofdiversities some set it would be in a rare gene family just affecting our visual acuity. It affects muchlife on the planet. We can line all these proteins; this was early on when we only had a couple ofthousand of them. And why do this?It turns out thereÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s a single amino acid residue that determines the wave length of light that thesereceptors see. So then that allows us to ask some unique questions. Do we see any association withdifferent geographic regions with the wavelength of light? And weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re quite surprised in fact to seesomething that makes sense. But nobody predicted ahead of time. For example, the Sargasso Sea, itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢sa deep indigo blue the organisms that are there the photoreceptors see blue light. You get into a coastalwaters where thereÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s a lot of chlorophyl, they see primarily green light. Yet in the fresh water like thePanama Canal they see entirely green light.So, thereÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s only one letter that genetic code then this needed to change to change the wavelength oflight by changing this amino acid residue. And even though there is a large abundance of organismstraversing these waters. The ones that survive and grow are the ones that can get their energy directly from the sun.And last year there was a study from the Swedish group showing in fact these organisms grow directlyon sunlight. This is not photosynthesis. This is the biological mechanisms that you are seeing in theslide with right now. You're having light hit these receptors and they transport ions across themembrane well in these microorganisms. ThatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s how they generate their energy in the low nutrientenvironment. So, in fact, instead, of low nutrients indicating and the scarcity of life this was the someof the highest density of life anywhere that weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ve seen because they have these mechanisms of gettingof getting energy straight from sunlightThe other thing that weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ve found is where people thought there was a single organism, we foundevidence for thousands. All related in the sense they have similar sets of genes, similar gene order, buthardly a single one. Each one of these little bars you see on here represents 900 base pairs of geneticcode. One of the things we can do is take a slice out of this date anywhere and create trees to look atthe relationships between these very closely related organisms. TheyÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re color coded by sights. So wecan look at things like Atlantic Ocean versus the Pacific Ocean. But the bottom to me is the mostexciting when you look at recent evolutions. ThereÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s been a switch between blue and green light forthese photoreceptors for different timesSo you can see at such selective pressure, you get a mutation and you switch from blue to green andyou're in coastal waters obviously you're going to thrive. If you switch fro blue to green out in themiddle of the ocean you probably won't. You can ask basically any question of this data in a similarfashion. ThereÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s in fact a lot questions. How much novelty is there with all these new discoveries thathave been doubling the number of all genes known to science. Are they like the photoreceptors justnew members of known gene families? Or are we really making new discoveries and if we are makingnew discoveries whatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s the pace of these discoveries?When we look at the dataÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s had initially weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re quite shocked to find that our database just betweenHalifax and the Galapagos was twice the size of all the public databases in terms of gene content. Andso we did a calculation by comparing all those data to each other. It was about a million CPU hoursand we found out thus far in the data set, there was maybe 50,000 major gene families. Well, we onlyhave 22 ÃƒÂ¢Ã¢â€šÂ¬Ã¢â‚¬Å“ 23,000 genes, maybe, 14 or 15 gene families so even all of human biology that representsonly a subset of this data but if we look at this as a cur instead of a long now, we have a long tale thatbasically goes out approaching infinity right now with new discoveries and new gene families with theexception of the animal world. So just to remind people, weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re just in one part of evolution eventhough the tree model doesn't really hold; it is useful for these kinds of diagrams.When we look at our reading the genetic code in mammals, it is basically been saturated so sequencinganother Mammalian Genome, your favorite pet, the species down the street, whatever, youÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re not goingto discover any new genes. You might discover some unique combinations or spawning of those genesbut we pretty much saturated the mammalian part of the tree but if we look at bacteria and archaeaweÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re in a linear phase of discovery. We donÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢t really know where we are, we just know itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s linear.Our thinking is that weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re in the earliest phases of this, just a few percent at most. Which meansanybody can just go out to the bay or go across to the Pacific, take a sample of seawater and make tensof thousands of species discoveries, millions of gene discoveries, et cera.We first, actually, applied these techniques to the human, looking at the bacteria that are associatedwith us. So the micro biome is your collection of microbes, not all of you have all these cavities, butyou know, this is no time for cavity jealousy or anything but we can isolate organisms from thesedifferent cavities and for example, look at the person next to you and you know, maybe you see themas having thousands of species in their mouth right now or maybe you can taste your own species.Only about 4 million genes of foreign species in your mouth, in your intestinal tract, if you have thoseother parts, you have those as well. So we have more bacterial cells associated with us than we havehuman cells. And it turns out; they affect our physiology in some pretty interesting ways.ThereÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s a company in North Carolina called Metabolomics that is using High-throughput MassSpectrometry to look at all the chemicals in the blood stream and theyÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ve worked out that we, as aspecies, can maybe make around 2,400 chemicals. If you look in the bloodstream of anybody, forexample, after theyÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ve had a meal, easily find about 60% of whatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s there is made by our own bodies.About 30% are just chemicals that came out of what we ate. You know, so this notion ÃƒÂ¢Ã¢â€šÂ¬Ã…â€œyou are whatyou eatÃƒÂ¢Ã¢â€šÂ¬ is partly right but 10% are bacterial metabolites so in fact, partly we are what we feed ourbacteria and what they give us.But weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re dealing at any one time with hundreds to thousands of foreign chemicals circulating in ourbloodstream, not to mention what we add directly by taking pharmaceutics or other things and we haveno idea, the impact of these on physiology because, in fact, we didnÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢t even know they were there beforeso weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re looking at the complexity of the human genome and all the variations there and whatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s in ourbloodstream. And if we did this test on everybody in this room, weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ll get a different answer foreverybody based on what you ate and also the uniqueness of the bacteria in your own guts, in yourmouth. This affects our chemical milieu. This is part of the environment that affects us.WeÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re also sequencing the air genome. So for example, in a room this size, sitting here for an hour,you would absorb maybe 10,000 different bacteria and maybe 10 times that in viruses in hour. If yougo outside, itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s twice that much so you're actually safer in here, right now, depending on who youÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢resitting next to, again, and what theyÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re exhaling and inhaling. So we live in a bacterial milieu. YouÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢regoing to the water; youÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re surrounded by bacteria so if youÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re out swimming and you swallow amouthful of seawater, you just swallowed millions and millions of bacteria and viruses. The air webreathe, the soil, our own skin, our own cavities so we are as much dependent on bacterial metabolismas human metabolism.WeÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ve been collecting all these genes, all these information, trying to understand the complexity. Andwe decided one way to try and understand the complexity was to try and mimic it. This is standard inchemistry to prove that you have the structure that you think you have. You remake that compound.We decided to try and do this with the genetic code starting with some pretty simple organismsThe second organism that we sequenced in 1995 was Mycoplasma genitalium. This is its genome, atleast, its depiction of it. Very much unlike our genome so it has a little over 500 genes in contrast toour 22,000. We have gaps in our genetic code with no genes that are much larger than this entiregenetic code. So bacterial genomes, you can see these bars represent genes, have very little intergenicspace. This is still the smallest genome of a self-replicating organism. So, theyÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ll grow on its own inthe lab. There are smaller genomes, but they're dependent on having a symbiote organism for itsgrowth. So, itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s not clear whether they're the extension of the viral world or just truly simple organismsthat are dependent on others. And we just ask simple questions, if one cell needs 18,000 genes to liveand this one needs 500 is this actually the minimal set. Can we define life in molecular terms based onthe genetic code?And so, we set out to try and knock out genes in this genome to see if we could get down to a smallernumber. So, every place you see one of these small triangles is where a transposon which is just apiece of DNA very much like a small virus than can randomly insert anywhere in the genetic in thegenetic code. Then we select for living cells. So, only genes that are not essential can toleratetransposons going into them. So, this is very much a negative map. You can see some genes can takelarge numbers of transposons, but keep in mind these experiments only done gene at a time. And wegot to a number of around a hundred genes that we could knock out. But we doubted whether if weknocked all 100 out that we could get to a living cell. In fact, if we look at a metabolic map of thissimplest of organisms it may look complex, but this is remarkably simple from looking at any one ofcells in the human body. And hereÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s all, if all the different genes that could be knocked out wouldknock out a lot metabolic pathways probably not resulting in a living cell.So, we decided the only way for it was to actually make chemically in the lab this chromosome. So wecould alter its gene composition and thatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s how the whole notion of synthetic genomics was born withthat relatively simple notion. So we started of with two primary questions, can the chemistry actuallypermit making these incredibly large biological molecules accurately in the lab and if we make it whatcan you do with it? DNA is an inner chemical, can we actually boot it up in to a living cell. This iscomparing the two areas. So the red line here is reading the genetic code and the blue line is our abilityto synthesize DNA. Its actually five orders of magnitude slower right now for writing the genetic codethan it is reading it. But this is changing exponentially and basically just over the last few months.And I think that writing the genetic code will soon catch up with our pace of reading it even though itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s still changing.Now, we thought this would be relatively simple. So, there are machines called DNA synthesizers thatcan make very short stretches of DNA called oligonucleotides. We make pieces that are about 50letters long and we thought if we just make a number of pieces and they overlap each other that theywould just all go together and we could make a larger piece of DNA. It turns out you can but theprocess with these machines for synthesizing DNA is very inaccurate. ItÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s a degenerative process, sothe longer you make the piece of DNA the more errors there are with it.So, we set out to do an experiment trying to make the Fi x 174 genome. Fi x 174 for those of you whodonÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢t know is a bacteriophage itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s a virus that kills bacteria particularly E. coli. It is one of the firstviruses that were extensively studied. In fact, it was the first actual genome of any type that was doneby Fred Sanger. We chose this because in fact its genome is very intolerant of changes. If you changethe genetic code the virus cannot reproduce. So, we developed some new techniques for actuallyaccurately writing the genetic coed where we can repair the errors in real time as were making it. Andfor us even though there had been an attempt by another group to make the polio virus which wasslightly larger than this. It only had one ten thousandth of the activity because of all these errors andmaking the genetic code. We went from the genetic code in the computer designing the pieces so thatthey would go together appropriately use this new process for error correction and we ended up with apiece of DNA exactly the right length. When we sequenced it was exactly what we had designed andwere trying to make.The exciting part was we took this piece of DNA and inserted into the bacteria E. coli and what hadhappened was E. coli recognized this as a piece of software and started making viral particles. Andtrue to form in nature when the viral particles were released from the cell. They turned around andkilled the bacteria that had made it. So, this is a process that we see all the time in nature. I was justspeaking to oil executives and I said they clearly understood that process. (Laughter) But this waspretty exciting of just taking a piece of DNA and having it activated making viral particles. So weview this as the software actually building its own hardware.This is an important concept as weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re trying to go forward in this field, that even most people that areworking in this area have not truly grasped the implications of this, that we donÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢t have to design lifefrom scratch. We just have to design the software appropriately. In fact weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ve gone to scale this up.Our plan was to make pieces that were viral-sized pieces that we would then out together to make anintact chromosome. We look for a number of ways to do this. We had been working on it for over 4years and thereÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s a process in nature called homologous recombination. This is that paper that youmight have read about that we just published where we in fact made this entire chromosome. ItÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s thelargest man-made molecule of a defined structure. To print it out, one letter at a time at 10 font with nospaces, it takes 142 pages just to print out the letter code for this structure. Its 582,970 base pairs orletters of genetic code and itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s over 300 million molecular weight. So, the process that we used wasnot as simple as just making pieces go together but itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s close to that. But it started with the rightdesign. And Design Now is a key part of biologyIn fact, before we started even the design phase we had to go back and re-sequence the entire genomebecause the standard in 1995 was roughly one error per 10,000 letters of genetic code. Nobody thoughtabout when reading the genetic code even with that seemingly accurate error that anybody would beusing those sequences to reproduce the organisms to actually write the genetic code. And if youÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢restarting with the genetic code of the computer, you can only make things as accurate as the informationyouÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re starting with. So, we went back and sequenced the genome all over again and found 30 errors,which was the standard for where it should have been. But we know with those 30 errors if we madethis chromosome based the initial sequence. It would never been able to be booted up.So we start with this digital information and we design the pieces thinking that weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re now this 580,000piece into 50 base pair of segments. The 50 base pair of segments have to be designed so they overlapwith our neighbor and we started off by making the pieces on the order of 5 to 7 kb. Five to seventhousand base pairs. That each had to be design so when they line up with their neighbor they wouldoverlap. We designed some other unique elements into it. You might be able to see this on the top linethat shows the places in the genome where we inserted watermarks. We wanted to absolutely be surethat we would not fool ourselves or others by having a contaminant of even one molecule of DNA fromthe native organism and this could be the fun part of this process. So, we have a four-letter genetic code.I'm sure most of you know that we have this triplet code were three letters of the genetic code, code forour amino acids. And we have roughly 20 amino acids and there's a single letter code representingeach amino acid. So, we can write things in the genetic code using this procedure. And we use this infact to label the DNA and the team came up with though some of the authors of the genetic code andthe institution. WeÃƒÂ¢Ã¢â€šÂ¬Ã‹Å“ve read that people were very disappointed. We didnÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢t put any poetry or othermore profound statements like one small step or anything like that. But weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ll think about that morecarefully for the next time. So, all this went into design started making these pieces, we put fourthese 5 to 7,000 base pair pieces together to make 24 kb pieces and at each stop we would grow thesepieces up in bacteria E. coli to make large amount of the DNA that we then sequence to make sure itwas absolutely accurate because not only were we trying to make the end product, which we couldhave done much faster. We were trying to make absolutely robust methods so we would understand iserrors crept in where they came fromWe combined the 24 kb pieces to make 72 kb pieces and the prior world record for the largest piece ofDNA made was around 31,000. So all these pieces at 72,000 greatly exceeded what had been donebefore. Then we put those together. This looks like a basketball playoff perhaps to make what wecalled quarter molecules and half molecules. But what we found is exceeded the limits of cloning andbacteria and we switched to another system. We switched to yeast to put these together and to growthem up. In fact, we had always envisioned that we put these pieces together by this process calledhomologous recombination.Basically, cells and nature do this all the time with repairing their DNA. In fact, this organismDeinococcus radiodurans which is pretty ubiquitous on the planet and perhaps elsewhere can take 3million rads of radiation. Its chromosomes get blown apart with a couple of hundred little pieces thanas long as itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s in water over 12 to 24 hours it remakes its chromosome exactly as it was before. Soheres a actual picture of it after 1.75 million rads of radiation. I recommend you not try at homebecause we as a species can only take a tiny, tiny fraction of this amount of radiation without beingkilled. We can't do this with our human chromosomes.So this is pretty stunning process it turns out this cell was not unique. We have thousands of species onthis planet that can do this. These cells can be totally desiccated. They can be dried out. They can bein a vacuum. They can accumulate this ionizing radiation for a very long period of time. We donÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢tactually know how long, but the speculation and calculations weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ve done is it would certainly fit in tothe Long Now Foundation because itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s tens of thousands to millions of years if the organisms even hadslight shielding in a comet or other material. We know this organism can survive in outer space. Itreaches an aqueous environment and reassembles its genetic code and it can start replicating again.So, we thought these mechanisms that weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re trying to isolate from this organism would be great forassembling the little pieces of the genome to put it back together again. But in fact, we found it waseven simpler than that. It turns out yeast, which is used for making bread and beer and wine all thesegood things we like can do this on its own with foreign pieces of DNA. So, while we are trying togrow up the pieces in yeast. We found that if we design the pieces correctly and out them in, it wouldassemble those automatically until the larger pieces into the intact chromosome. So thatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s how weended up with the entire 580,000 base pair piece that we sequenced and itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s down to 0 errors. This isactually a picture of it. You donÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢t need an electron microscope because this molecule is so large. Thisis just looking at over a 6 second period as you can actually see itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s a circular piece of DNA. ItÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s prettyexciting for us to be able to actually visualize it.Well, how do you boot up a chromosome? You saw with the virus all we had to do was insert it in abacteria and the bacteria can start reading that software and producing things. We think itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s a little bitmore complicated with whole bacterial chromosomes. Initially, we thought that we would have to tryand remove the chromosome from a bacterial cell and add in this new to replace it. It turns out thatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢svery hard to do. So what youÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ve heard about with mammalian cloning works very easily in eukaryoteswhere thereÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s a defined nucleus thatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s easy to just to cut out of the cell under a microscope. Lift it outand out in a nucleus from a different organism or a different cell from the same organism.With bacteria and archaea, there is no nucleus. The DNA is part of the cytoplasm of the cell so wehave to use a little be more ingenuity to do this and last year we published what we think is the keytechnique of transplantation thatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s pretty stunning in its own right because we, actually, by putting in anew chromosome into a cell that had a different chromosome in it to start with. We completelyconverted one species into another so it was an absolute 100% transformation. We used two closelyrelated mycoplasma species roughly the same distance apart as we are to mice. We isolated thechromosome from this M. mycoides cell and we wanted to make sure we can get down to justDNA. Chromosomes have a lot of protein associated with them so we treated these pretty harshly withdigestive enzymes to digest away all the proteins. And then we added back a few additional genes tothis chromosome. One set of genes, the lacZ set, for example, will turn the cells blue so it makes iteasy to identify them. And we also added in a set of selectable markers so we could select for cells justfor this transplanted chromosome and we put this chromosome into a species called a capricolum.Now, hereÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s a wonderful graphic. I'm sure you all appreciate the sophistication of this. Wetransplanted the chromosome into the cell. In fact, we thought we would end up with this situation, acell with two chromosomes in it. We see this all the time in nature, so all these people that makearguments against evolution because we know that you can't just get a point mutation in one piece ofDNA unless itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s for the wavelength of light your photoreceptor sees but thatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s going to change intomore complexity. What we see in the real world is we see chromosomes moving around where youcan add a thousand new traits to a cell in a second.Cholera, for example, most people thought there was no point in decoding the cholera genome becauseit was very closely to E. coli but when we read the cholera genome it turns out that it didnÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢t have just asingle chromosome. It had two chromosomes and it looked like this. They were very different fromeach other. Obviously, it had taken up a chromosome from another species and added it to itsrepertoire. But we didnÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢t want this situation. It turns out the capricolum genome is very unusual inthat it doesn't contain any genes for restriction enzymes. Restriction endonucleases are the molecularscissors that cut up DNA and it is how cells protect themselves from this foreign DNA coming in. Infact, the chromosome that we put in did have a restriction enzyme. As soon as it was in the cell, it gotexpressed and it recognized the original chromosome as foreign and cut it up into small pieces and itgot digested. So were left with cells just with the transplanted chromosomesIn over a very short period of time, we ended up with these blue cells that all the characteristics of thesecells were that what was dictated by the transplanted chromosome. Every protein in the cell changedfrom that with the original species into that coded by the M. mycoides chromosome, the membranechanged, everything changed. We can isolate the DNA and it was only what we had transplanted in.So this is true identity theft at the ultimate level and fortunately, either most of us have mechanisms toprotect us against this, and most cells do with these restriction enzymes but they give us very powerfultools to try and do this in the future so we know we can take a chromosome. We can transplant it. Wecan completely convert one species into another.So weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re in the process of doing this right now with the synthetically made chromosome. We hope thiswill happen very soon. These experiments go very slowly because the cells grow slowly. So it takesabout 6 weeks once you do a transplant to see whether you have viable cells but we should be seeingthat, hopefully, in the very near future. That will complete the trilogy that we had been putting togetheron this, although, itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s clear now just from the transplant experiment that we know this will work and tome, itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s more of a technicality if it works with synthetically-made DNA versus DNA out of a cellbecause it is the same ÃƒÂ¢Ã¢â€šÂ¬Ã¢â‚¬Å“ it sequences the same but I think itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s still important for the proof. But ittransforms us into a new era of now (indiscernible)[00:51:47] alter cells by simply rewriting the genetic code.So the next steps and why. I'm going to walk you through quickly. Some of the reasons why weredoing this and where were trying to go. But a lot of people when they think about this and write aboutit, they think weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re redoing Genesis and weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re not. This is much more like a new version of theCambrian explosion. WeÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re dependent as the title of this lecture said on the 3 1/2 billion years ofevolution. Also in because weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re as humans because of that and weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re using organisms that haveevolved and substantial period of time. WhatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s different is that we donÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢t have to redo all of thatbecause if we just write new molecular software, we can start at that point and go in an infinite numberof directions. Why do this? One of the reasons is trying to deal what the future of our species on thisplanet over the next 40 years weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re going to go from 6 1/2 to 9 billion people. ThatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s a huge change ina short period of time. I try to put this data in context for myself and hope that it leads others to be able to understand it.So I was born in 1946. There's now three people on this planet for everybody that existed in 1946. Soif you think of having 1/3 of the number of people in this room in 40 years of before people foreverybody that was on the planet in 1946. WeÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re having trouble providing food, clean water, energy,housing for the 6 1/2 billion. WeÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re going to have even more trouble trying to provide it for 9 billionpeople. WeÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re changing our environment quite dramatically by burning billions of years of biology inthe form of oil and coal. This is now from 2003 so, you know, over 5 billion tons of coal. Billions ofbarrels of oil that we just take out and burn and the CO2 goes into the atmosphere. The slide is out ofdate. These should be changed again the third time in a year. This number is now 4.2 billion tons ofCO2 that were adding and stays in the atmosphere each year. That number is accelerating. The oceansink which is the largest sink as you can see that they can take hundred of billions of tons of CO2.Many people think itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s saturated and deforestation is also contributing as we try and strip more land tomake more food and fuel everybody who have seen this graph and the steady increase of CO2. So what can be done?There is a dramatic revolution thatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s taking place in the industrial world equivalent maybe to the firstindustrial revolution. Companies like DuPont which was basically built on using oil as its rawmaterial, had now had to switch away from using oil. They are switching to sugar which maybe wonÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢tbe anymore sustainable and well talk about that in a second. But they spent 10 years but over 100million dollars offering E. coli to do a simple reaction. To make 6carbon sugar and make 3 carbonpropane dial molecule. And if you can see those what looks like a large beer kegs, those are actually 46,000 liter fermenters that they grow up literally 100 tons of this bacteria at a time in a batch and makepropane dial from sugar with this engineered bacteria. They can do this cheaper and faster than theycan with chemical conversion with their chemists.This has certain downfalls after each batch. They have to bury those tons of E. coli in a land fillbecause they are not allowed to burn it. But even all these limitations and the cost of sugar they claimthat their new polymer that uses this chemical will be the first multi-billion dollar biotech productthatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s not a pharmaceutical. Some metabolic engineering is being used. My new company Syntheticgenomics has a deal with VP to try and use biology deep in the Earth to stop mining coal bybiologically that coal into methane. We were quite surprised, a mile down in the Earth when we tookthe first sample. There was more biological diversity there than we found in the ocean. Just a simplemicroscopic field was teeming with microorganisms. This is a piece of coal that we have in laboratory.All those little tiny things or the bacteria that live off a coal was substrate and we have other bacteriathat convert that substrate right into methane. ItÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s a little bit dark but maybe you can see the methane.Bubbles coming off a coal in the bottom.This doesnÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢t stop taking carbon under the ground but itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s about a 10 fold improvement over miningcold and burning it. And the coal reserves are extensive enough that this could maybe get ourselves astopgap until there is a new economy. But weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ve been going in the wrong direction already and partbecause of lobbies in the government corn to ethanol. Just does not going to get us there, itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s anegative carbon balance itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s been heavily subsidized by all of us. And the numbers are pretty stunningit looks like a huge amount of ethanol were being produced and we actually look at the numbers on theorder of 6 billion gallons. There's now I think 160 plants around the country, 6 billion sounds like a lotuntil you look at what we use. So this is our transportation fuel, 140 billion gallons of gasoline, 45billion gallons of diesel. So 5 to 6 billion gallons of ethanol basically does nothing for this equation.Especially when it produces more CO2 than it captures.ItÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s competing with farmland. Food prices are going up because of this. Corn prices have doubled onthe last year. So this is just the wrong experiment taking us very much in the wrong direction. Now,were not short of energy on this planet. A hundred twenty thousand terawatts of energy arrives hereeach day from the sun. If we could capture that at 1% efficiency, weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢d only need about 5 billionhectares. IÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ve argued this is a great use of Nevada. Nobody has ever argued with it. They havenÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢tcome up with better alternative views. We could just from sunlight produce energy. In fact, desertsand seawater donÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢t compete with food. WeÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re in fact working on, IÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ll show you in a minute what werecalling a fourth generation fuel starting with CO2 as the feed stock. So if we can start with CO2 eitherfrom sunlight in the methanogenesis pathways. We skip all this use of farmland for producing fuels.We have two fuels that are pretty close going into test production we call them second and thirdgeneration. They both have the downfall of using sugar as a starting material but they're far superior toethanol. They donÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢t mix with water. They have very low freezing points they can just be fluffed andblended they can go through pipelines they have a much higher energy density to get people going inthe different direction, but the one that we think weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ll have in about 18 months, the fourth generationfuels, starts with CO2 as a feed stalk. Now, if we can really use CO2 as a feed stalk it does severalthings when you consider the efforts for CO2 sequestration. We can just take piped CO2 into reactors,biological reactors and make fuel such as octane directly from carbon dioxide, either using sunlight asthe energy source or molecular hydrogen.You wonÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢t see any energy plans incorporating this types of ideas. WeÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢d go forward by extrapolatingfrom the past from linear ideas or linear thinking. There is no planning for disruptive technologies, butin fact if were going to change in anything in anybodyÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s lifetime here, we need disruptive technologiesat the Davos meeting this year the conclusion was 40 years from now biology and all alternate energysources will maybe only make a few percent impact on using oil and coal, because there's so muchvested interest. There's a new cold powered fuel power plant coming online everyday. These causedbillions of dollars to build. So, itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s only if thereÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s an economically viable alternative that doesnÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢tcompete with food that has a chance to change anything in the near future and I donÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢t think we want tokeep adding CO2 to the atmosphere.So we have 10 million genes in the databases right now. WeÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re going to double that number again thisyear from the Sorcerer expedition to 20 million. These are our design components. The entireelectronic industry only had a few handfuls of designed components and look at the diversity that wehave there. We have tens of millions of starting components that we can help. And now we built arobot that instead of making one chromosome per 4 years. We think we can get up to makingthousands to millions to millions of them a day and in random combinations. So were calling this newfield combinatorial genomics. So just think of it, if we can basically start with design take these tens ofmillions of design components. Just make the DNA transform that automatically in to yeast or someother vector and screen for chemical production for octane production antibody production. Whateveryou screen for will be the selection that happens. It will change the pace of biology in unimaginableways. Just like I donÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢t think anybody could imagine in a 1950s this laptop computer went close toterabyte of storage on it doing more than giant rooms could. This is a new area thatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s going to bedriven by peopleÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s imagination. This is sort of software that were working on. To make the newsoftware thinking of this as a true design phase. Trying to come up with new fuels, new chemicals.And so itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s an exciting time as we try to scale this up with remarkably small teams. This is a differentaspect of industrial revolution. And that it doesnÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢t take armies of thousands.In fact thatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s what weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ve done from the beginning with reading the genetic code. The yeast genometook 10 years and a thousand scientists. We reduced that project first to 4 months and now to ours andwere doing the same thing now going in the other direction. ItÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s clear weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ll be able to have a huge dentin the first bullet here in increasing the understanding of life because we need to use empirical methodswith the huge data sets that we have. There's not enough scientists to ask reasonable questions to workout the biology of whatÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s been discovered to date. But IÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢m more optimistic I'm hoping within 15-20years we can really have gone a long way for replacing the petrochemical industry. Hopefully, biologywould be major source of energy. And bioremediation if we can capture back CO2 and recycle thatinto fuel. We possible even start to undo some of the damage that weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ve been doing for the last 100years. And were also looking at this in terms of every type of chemical and looking at making forexample combinatorial anti-bodies so the we can have instant vaccines for every variant of the fluvirus for example instead of being a year late like we are again this year. This field is unusual and thisis my last slide by the way. It started with us asking the ethical questions before we did the firstexperiment not after we thought ÃƒÂ¢Ã¢â€šÂ¬Ã…â€œGee whiz, we have a lot of potential here.ÃƒÂ¢Ã¢â€šÂ¬ The result of this team atthe University of Pennsylvania published their review in 1999. They thought we were proceeding withthe right questions and the right technology. There was a caveat about being concerned aboutbiological warfare.The Sloan Foundation just funded my institute and MIT for the last couple of years to do a completereview of the risks and the benefits. I think I published at the end of last year and there are ongoingdiscussions a new body in the federal government got formed when we published Fi X 174 genome.Got reviewed at the White House, they had to decide whether to try and make our data and processessecret. I think its one of the few good things of the Bush administration has done in sciences. Theywent and opted for open publication. But they formed a new branch, executive committee that haspeople from every branch of the government reviewing this type work weÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢ve had public discussionsongoing basically since 1996 on this. WeÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢re trying to take it one step at a time but itÃƒÂ¢Ã¢â€šÂ¬Ã¢â€žÂ¢s about to expandin an exponential fashion and I think its going to be an exciting next 10,000 thousand years.Thank you very much.