Kary Mullis Science, like nothing else among the institutions of mankind, grows like a weed every year. Art is subject to arbitrary fashion, religion is inwardly focused and driven only to sustain itself, law shuttles between freeing us and enslaving us Kary Banks Mullis (born December 28, 1944) is a Nobel Prize winning American biochemist, author, and lecturer. In recognition of his improvement of the polymerase chain reaction (PCR) technique, he shared the 1993 Nobel Prize in Chemistry with Michael Smith[1] and earned the Japan Prize in the same year. The process was first described by Kjell Kleppe and 1968 Nobel laureate H. Gobind Khorana, and allows the amplification of specific DNA sequences.[2][3] The improvements made by Mullis allowed PCR to become a central technique in biochemistry and molecular biology, described by The New York Times as "highly original and significant, virtually dividing biology into the two epochs of before P.C.R. and after P.C.R."[4] Since winning the Nobel Prize, Mullis has been criticized in The New York Times for promoting ideas in areas in which he has no expertise.[5] He has promoted AIDS denialism,[6][7][8][9][10][11] climate change denial[6] and his belief in astrology.[5][6] Early life and education Mullis was born in Lenoir, North Carolina, near the Blue Ridge Mountains,[12] on December 28, 1944. His family had a background in farming in this rural area. As a child, Mullis recalls, he was interested in observing organisms in the countryside.[3] He grew up in Columbia, South Carolina,[3] where he attended Dreher High School. Mullis earned a Bachelor of Science degree in chemistry[12] from the Georgia Institute of Technology in Atlanta in 1966, during which time he got married and started a business.[13] He then received a Ph.D. in biochemistry from the University of California, Berkeley in 1972; his research focused on synthesis and structure of proteins.[3] Following his graduation, Mullis became a postdoctoral fellow in pediatric cardiology at the University of Kansas Medical School, going on to complete two years of postdoctoral work in pharmaceutical chemistry at the University of California, San Francisco. Career After receiving his PhD, Mullis left science to write fiction, but quit and became a biochemist at a medical school in Kansas City.[13] He then managed a bakery for two years.[4] Mullis returned to science at the encouragement of friend Thomas White, who later got Mullis a job with the biotechnology company Cetus Corporation of Emeryville, California.[3][4] Mullis worked as a DNA chemist at Cetus for seven years; it was there, in 1983, that Mullis invented his prize-winning improvements to the polymerase chain reaction.[14] After leaving Cetus in 1986, Mullis served as director of molecular biology for Xytronyx, Inc. in San Diego for two years. Mullis has consulted on nucleic acid chemistry for multiple corporations.[4] In 1992, Mullis founded a business with the intent to sell pieces of jewelry containing the amplified DNA of deceased famous people like Elvis Presley and Marilyn Monroe.[15][16] Mullis is also a member of the USA Science and Engineering Festival's Advisory Board.[17] Books authored The Polymerase Chain Reaction, 1994, with Richard A. Gibbs Dancing Naked in the Mind Field. 1998, Vintage Books. Mullis's 1998 autobiography Dancing Naked in the Mind Field, gives his account of the commercial development of PCR, as well as providing insights into his opinions and experiences. In the book, Mullis chronicles his romantic relationships, use of LSD, synthesis and self-testing of novel psychoactive substances, belief in astrology and an encounter with an extraterrestrial in the form of a fluorescent raccoon. Awards and honors 1990 William Allan Memorial Award of the American Society of Human Genetics | Preis Biochemische Analytik of the German Society of Clinical Chemistry and Boehringer Mannheim 1991 National Biotechnology Award | Gairdner Award | R&D Scientist of the Year 1992 California Scientist of the Year Award 1993 Nobel Prize in Chemistry | Japan Prize | Thomas A. Edison Award 1994 Honorary degree of Doctor of Science from the University of South Carolina 1998 Inducted into the National Inventors Hall of Fame [33] | Ronald H. Brown American Innovator Award[34] 2004 Honorary degree in Pharmaceutical Biotechnology from the University of Bologna, Italy Mullis also received the John Scott Award in 1991, given by the City Trusts of Philadelphia to others including Thomas Edison and the Wright Brothers.[35] PCR and other inventions Main articles: Taq Polymerase and History of polymerase chain reaction In 1983, Mullis was working for Cetus Corp. as a chemist.[13] That spring, according to Mullis, he was driving his vehicle late one night with his girlfriend, who was also a chemist at Cetus, when he had the idea to use a pair of primers to bracket the desired DNA sequence and to copy it using DNA polymerase, a technique which would allow a small strand of DNA to be copied almost an infinite number of times.[13] Cetus took Mullis off his usual projects to concentrate on PCR full-time.[13] Mullis succeeded on demonstrating PCR December 16, 1983.[13] In his Nobel Prize lecture, he remarked that the success didn't make up for his girlfriend breaking up with him shortly before: "I was sagging as I walked out to my little silver Honda Civic. Neither [assistant] Fred, empty Beck's bottles, nor the sweet smell of the dawn of the age of PCR could replace Jenny. I was lonesome."[13] He received a $10,000 bonus from Cetus for the invention.[13] Other Cetus scientists, including Randall Saiki and Henry Erlich, were placed on PCR projects to work on developing HIV- and other tests utilizing PCR. Saiki generated the needed data and authored the first paper to include utilization of the technique,[4] while Mullis was still working on a paper that would describe PCR itself.[13] A complication at that point was that the DNA polymerase used was destroyed by the high heat used at the start of each replication cycle and had to be replaced. In 1986, Mullis started to use Thermophilus aquaticus (Taq) DNA polymerase to amplify segments of DNA. The Taq polymerase was heat resistant and would only need to be added once, thus making the technique dramatically more affordable and subject to automation. This has created revolutions in biochemistry, molecular biology, genetics, medicine and forensics. Mullis has also invented a UV-sensitive plastic that changes color in response to light, and most recently has been working on an approach for mobilizing the immune system to neutralize invading pathogens and toxins, leading to the formation of his current venture, Altermune LLC. Mullis described this idea this way: It is a method using specific synthetic chemical linkers to divert an immune response from its nominal target to something completely different which you would right now like to be temporarily immune to. Let's say you just got exposed to a new strain of the flu. You're already immune to alpha-1,3-galactosyl-galactose bonds. All humans are. Why not divert a fraction of those antibodies to the influenza strain you just picked up? A chemical linker synthesized with an alpha-1,3-gal-gal bond on one end and a DNA aptamer devised to bind specifically to the strain of influenza you have on the other end will link anti-alpha-Gal antibodies to the influenza virus and presto!--you have fooled your immune system into attacking the new virus.[12][18] Medical Applications PCR has been applied to a large number of medical procedures: The first application of PCR[1] was for genetic testing, where a sample of DNA is analyzed for the presence of genetic disease mutations. Prospective parents can be tested for being genetic carriers, or their children might be tested for actually being affected by a disease. DNA samples for Prenatal testing can be obtained by amniocentesis, chorionic villus sampling, or even by the analysis of rare fetal cells circulating in the mother's bloodstream. PCR analysis is also essential to Preimplantation genetic diagnosis, where individual cells of a developing embryo are tested for mutations. PCR can also be used as part of a sensitive test for tissue typing, vital to organ transplantation. As of 2008, there is even a proposal to replace the traditional antibody-based tests for blood type with PCR-based tests.[2] Many forms of cancer involve alterations to oncogenes. By using PCR-based tests to study these mutations, therapy regimens can sometimes be individually customized to a patient. Infectious disease applications Characterization and detection of infectious disease organisms have been revolutionized by PCR: The Human Immunodeficiency Virus (or HIV), responsible for AIDS, is a difficult target to find and eradicate. The earliest tests for infection relied on the presence of antibodies to the virus circulating in the bloodstream. However, antibodies don't appear until many weeks after infection, maternal antibodies mask the infection of a newborn, and therapeutic agents to fight the infection don't affect the antibodies. PCR tests have been developed that can detect as little as one viral genome among the DNA of over 50,000 host cells.[3] Infections can be detected earlier, donated blood can be screened directly for the virus, newborns can be immediately tested for infection, and the effects of antiviral treatments can be quantified. Some disease organisms, such as that for Tuberculosis, are difficult to sample from patients and slow to be grown in the laboratory. PCR-based tests have allowed detection of small numbers of disease organisms (both live or dead), in convenient samples. Detailed genetic analysis can also be used to detect antibiotic resistance, allowing immediate and effective therapy. The effects of therapy can also be immediately evaluated. The spread of a disease organism through populations of domestic or wild animals can be monitored by PCR testing. In many cases, the appearance of new virulent sub-types can be detected and monitored. The sub-types of an organism that were responsible for earlier epidemics can also be determined by PCR analysis. Forensic applications The development of PCR-based genetic (or DNA) fingerprinting protocols has seen widespread application in forensics: In its most discriminating form, Genetic fingerprinting can uniquely discriminate any one person from the entire population of the world. Minute samples of DNA can be isolated from a crime scene, and compared to that from suspects, or from a DNA database of earlier evidence or convicts. Simpler versions of these tests are often used to rapidly rule out suspects during a criminal investigation. Evidence from decades-old crimes can be tested, confirming or exonerating the people originally convicted. Less discriminating forms of DNA fingerprinting can help in Parental testing, where an individual is matched with their close relatives. DNA from unidentified human remains can be tested, and compared with that from possible parents, siblings, or children. Similar testing can be used to confirm the biological parents of an adopted (or kidnapped) child. The actual biological father of a newborn can also be confirmed (or ruled out). Research applications PCR has been applied to many areas of research in molecular genetics: PCR allows rapid production of short pieces of DNA, even when nothing more than the sequence of the two primers is known. This ability of PCR augments many methods, such as generating hybridization probes for Southern or northern blot hybridization. PCR supplies these techniques with large amounts of pure DNA, sometimes as a single strand, enabling analysis even from very small amounts of starting material. The task of DNA sequencing can also be assisted by PCR. Known segments of DNA can easily be produced from a patient with a genetic disease mutation. Modifications to the amplification technique can extract segments from a completely unknown genome, or can generate just a single strand of an area of interest. PCR has numerous applications to the more traditional process of DNA cloning. It can extract segments for insertion into a vector from a larger genome, which may be only available in small quantities. Using a single set of 'vector primers', it can also analyze or extract fragments that have already been inserted into vectors. Some alterations to the PCR protocol can generate mutations (general or site-directed) of an inserted fragment. Sequence-tagged sites is a process where PCR is used as an indicator that a particular segment of a genome is present in a particular clone. The Human Genome Project found this application vital to mapping the cosmid clones they were sequencing, and to coordinating the results from different laboratories. An exciting application of PCR is the phylogenic analysis of DNA from ancient sources, such as that found in the recovered bones of Neanderthals, or from frozen tissues of Mammoths. In some cases the highly degraded DNA from these sources might be reassembled during the early stages of amplification. A common application of PCR is the study of patterns of gene expression. Tissues (or even individual cells) can be analyzed at different stages to see which genes have become active, or which have been switched off. This application can also use Q-PCR to quantitate the actual levels of expression The ability of PCR to simultaneously amplify several loci from individual sperm[4] has greatly enhanced the more traditional task of genetic mapping by studying chromosomal crossovers after meiosis. Rare crossover events between very close loci have been directly observed by analyzing thousands of individual sperms. Similarly, unusual deletions, insertions, translocations, or inversions can be analyzed, all without having to wait (or pay for) the long and laborious processes of fertilization, embryogenesis, etc. Others PCR is also important in answering basic scientific questions. In the field of evolutionary biology, PCR has been used to establish relationships among species. In anthropology, it has used to understand ancient human migration patterns. In archaeology, it has been used to help identify ancient human remains. Paleontologists have used PCR to amplify DNA from extinct insects preserved in amber for 20 million years. The Human Genome Project, which had a goal of determining the sequence of the 3 billion base pairs in the human genome, relied heavily on PCR. The genes responsible for a variety of human diseases have been identified using PCR. For example, a PCR technique called multiplex PCR identifies a mutation in a gene in boys suffering from Duchenne muscular dystrophy. PCR can also be used to search for DNA from foreign organisms such as viruses or bacteria. References 1. ^ Shampo, M. A.; Kyle, R. A. (2002). "Kary B. Mullis--Nobel Laureate for procedure to replicate DNA". Mayo Clinic proceedings. Mayo Clinic 77 (7): 606. PMID 12108595. edit 2. ^ Saiki, R.; Gelfand, D.; Stoffel, S.; Scharf, S.; Higuchi, R.; Horn, G.; Mullis, K.; Erlich, H. (1988). "Primerdirected enzymatic amplification of DNA with a thermostable DNA polymerase". Science239 (4839): 487± 491. doi:10.1126/science.2448875. PMID 2448875. edit 3. ^ a b c d e 'Biotechnology 101'' by Brian Robert Shmaefsky. Books.google.com. 2006-10- 30.ISBN 9780313335280. Retrieved 2010-07-27. 4. ^ a b c d e f "Scientist at Work/Kary Mullis; After the 'Eureka', a Nobelist Drops Out" Nicholas Wade, The New York Times, September 15, 1998. 5. ^ a b c Johnson, G (2007-10-28). "Bright Scientists, Dim Notions". The New York Times. Retrieved 2010-08- 06. 6. ^ a b c d Mullis, K (1998). Dancing Naked in the Mind Field. Vintage Books. pp. 115±118, 143± 153. ISBN 0679442553. 7. 8. 9. ^ a b Thomas, Charles A. (1994). "''Reason'', June 1994". Findarticles.com. Retrieved 2010-07-27. ^ a b "''Washington Informer'', May 31, 2000". Highbeam.com. 2000-05-31. Retrieved 2010-07-27. ^ a b Kalichman, Seth (2009). Denying AIDS: Conspiracy Theories, Pseudoscience, and Human Tragedy. New York: Copernicus Books (Springer Science+Business Media). pp. 177±178.ISBN 978-0-387-79475-4. 10. ^ a b Maggiore C (2006). What If Everything You Thought You Knew About AIDS Was Wrong?. American Foundation For AIDS Alternative. ISBN 0-9674153-2-2. 11. ^ a b Nattrass, N (2007). "AIDS Denialism vs. Science". Skeptical Inquirer 31 (5). 12. ^ a b c "Official Nobel Autobiography". Nobelprize.org. 1998-03-21. Retrieved 2010-07-27. 13. ^ a b c d e f g hi j k l m n o p Yoffe, Emily Emily Yoffe (Vol 122, no. 1 (July) 1994: 68±75). "Is Kary Mullis God? Nobel Prize winner's new life". Esquire. 2. ^ "''The Economist'', 2004". Economist.com. Retrieved 2010-07-27. 3. ^ a b ''Life on the Edge: Amazing Creatures Thriving in Extreme Environments'' by Michael Gross. Books.google.com. 2001-01-24. ISBN 9780738204451. Retrieved 2010-07-27. 4. 5. 6. 7. ^ "''The Hastings Center Report'', 1998". Questia.com. Retrieved 2010-07-27. ^ "Advisors". Usasciencefestival.org. Retrieved 2010-07-27. ^ "Kary Mullis' next-gen cure for killer infections | Video on". Ted.com. Retrieved 2010-07-27. ^ ''Artificial DNA: Methods and Applications'' by Yury E. Khudyakov, Howard A. Fields. Books.google.com. 2003. ISBN 9780849314261. Retrieved 2010-07-27. 8. ^ Richard Bilsker. "Ethnography of a Nobel Prize". Hyle.org. Retrieved 2010-07-27. 1. ^ Fridell R (2005). Decoding life: unraveling the mysteries of the genome. Minneapolis: Lerner Publications. pp. 88. ISBN 0-8225-1196-7. 2. ^ "Confronting AIDS: Update 1988". Institute of Medicine of the U.S. National Academy of Sciences. 1988. "«the evidence that HIV causes AIDS is scientifically conclusive." 3. ^ "The Evidence that HIV Causes AIDS". National Institute of Allergy and Infectious Disease. 2009-09-04. Retrieved 2009-10-14. [dead link] 4. ^ Oreskes, Naomi (December 2004). "BEYOND THE IVORY TOWER: The Scientific Consensus on Climate Change". Science 306 (5702): 1686. doi:10.1126/science.1103618.PMID 15576594. "Such statements suggest that there might be substantive disagreement in the scientific community about the reality of anthropogenic climate change. This is not the case. [...] Politicians, economists, journalists, and others may have the impression of confusion, disagreement, or discord among climate scientists, but that impression is incorrect." 5. ^ "Joint Science Academies' Statement" (pdf). United States National Academies. 2005-07-06. Retrieved 2011-06-09. 6. ^ "Understanding and Responding to Climate Change" (pdf). United States National Academies. 2008. Retrieved 2011-06-09. 7. ^ "Christine Maggiore, vocal skeptic of AIDS research, dies at 52". Los Angeles Times. 2008-12-30. Retrieved 2008-12-30. 8. ^ Schoch, Russell (September 1994). "Q&A - A Conversation with Kerry Mullis". California Monthly (Berkeley, CA: California Alumni Association) 105 (1): 20. Retrieved 2008-03-11. 9. ^ Ann Harrison (2006-01-16). "LSD: The Geek's Wonder Drug?". Wired. Wired. Retrieved 2008-03-11. "Like Herbert, many scientists and engineers also report heightened states of creativity while using LSD. During a press conference on Friday, Hofmann revealed that he was told by Nobel-prize-winning chemist Kary Mullis that LSD had helped him develop the polymerase chain reaction that helps amplify specific DNA sequences." 10. ^ "BBC Horizon - Psychedelic Science - DMT, LSD, Ibogaine - Part 5". BBC. 1997. Retrieved 2009-10-16. 11. ^ http://www.lrb.co.uk/v33/n22/jenny-diski/what-might-they-want Review of "The Myth and Mystery of UFOs", by Thomas Bullard 12. ^ Golden, Frederic (2000-12-13). "''Time Magazine'', December 13, 2000". Time.com. Retrieved 2010-0727. 13. ^ "Hall of Fame/Inventor Profile". Invent.org. 1944-12-28. Retrieved 2010-07-27. 14. ^ "Nobel Prize Winner Among Rondal H. Brown Award Recipients". Uspto.gov. 1998-10-13. Retrieved 2010-07-27. 15. ^ "John Scott Award Winners". Garfield.library.upenn.edu. 2005-10-28. Retrieved 2010-07-27. PCR Application References 1. ^ Saiki RK et al. "Enzymatic Amplification of -globin Genomic Sequences and Restriction Site Analysis for Diagnosis of Sickle Cell Anemia" Science vol. 230 pp. 1350-54 (1985). 2. ^ Quill E "Blood-Matching Goes Genetic" Science Magazine (14 March 2008) pp. 1478-1479. 3. ^ Kwok S et al. "Identification of HIV sequences by using in vitro enzymatic amplification and oligomer cleavage detection." J. Virol. vol. 61(5) pp. 1690-4 (1987). 4. ^ Boehnke M et al. "Fine-structure genetic mapping of human chromosomes using the polymerase chain reaction on single sperm." Am J Hum Genet vol. 45(1) pp. 21-32 (1989).