Editing Kary Mullis (section)

== Career ==

After receiving his doctorate, Mullis briefly left science to write fiction before accepting the University of Kansas fellowship.<ref name="Yoffe 1994" /> During his postdoctoral work, he managed a bakery for two years.<ref name="Wade 1998">{{Cite news |last=Wade |first=Nicholas |title=Scientist at Work/Kary Mullis; After the 'Eureka', a Nobelist Drops Out |date=September 15, 1998 |work=The New York Times |department=Science Times |pages=F1, F5 |issn=0362-4331 |url=https://query.nytimes.com/gst/fullpage.html?res=9802E7DD1331F936A2575AC0A96E958260 |url-access=limited}}</ref> Mullis returned to science at the encouragement of UC Berkeley friend and colleague Thomas White, who secured Mullis's UCSF position and later helped Mullis land a position with the biotechnology company [[Cetus Corporation]] of [[Emeryville, California]].<ref name="Shmaefsky 2006" /><ref name="Wade 1998" /> Despite little experience in [[molecular biology]], Mullis worked as a [[DNA]] [[chemist]] at Cetus for seven years, ultimately serving as head of the DNA synthesis lab under White, then the firm's director of molecular and biological research; it was there, in 1983, that Mullis invented the [[polymerase chain reaction]] (PCR) procedure.<ref name="DNA's detective story">{{Cite news |author1=<!--Staff writer; no byline--> |url=https://www.economist.com/technology-quarterly/2004/03/13/dnas-detective-story |title=DNA's detective story |newspaper=The Economist |series=Technology Quarterly |date=March 13, 2004 |issn=0013-0613 |url-access=limited}}</ref>

Mullis acquired a reputation for erratic behavior at Cetus, once threatening to bring a gun to work; he also engaged in "public lovers' quarrels" with his then-girlfriend (a fellow chemist at the company) and "nearly came to blows with another scientist" at a staff party, according to ''[[California Magazine]]''.{{r|McDonald}} White recalled: "It definitely put me in a tough spot. His behavior was so outrageous that the other scientists thought that the only reason I didn't fire him outright was that he was a friend of mine."<ref name="McDonald"/>

After resigning from Cetus in 1986, Mullis served as director of molecular biology for Xytronyx, Inc. in [[San Diego]] for two years. While inventing a UV-sensitive ink at Xytronyx, he became skeptical of the existence of the [[ozone hole]].{{Citation needed|date=January 2024}}

Thereafter, Mullis worked intermittently as a consultant for multiple corporations and institutions on nucleic acid chemistry and as an [[expert witness]] specializing in [[DNA profiling]].<ref name=":2" /><ref name="Wade 1998" /> In 1992, Mullis founded a business to sell pieces of jewelry containing the amplified DNA of deceased famous people such as [[Elvis Presley]] and [[Marilyn Monroe]].<ref name="Gross 2001" /><ref name="Nelkin 1998">{{Cite journal |title=Homo Economicus: Commercialization of Body Tissue in the Age of Biotechnology |journal=The Hastings Center Report |last2=Andrews |first2=Lori |last1=Nelkin |first1=Dorothy |volume=28 |issue=5 |date=September–October 1998 |pages=30–39 |doi=10.2307/3528230 |jstor=3528230 |pmid=11656768 |issn=1552-146X}}</ref> In the same year, he also founded Atomic Tags in [[La Jolla, California]]. The venture sought to develop technology using atomic-force microscopy and bar-coded antibodies tagged with heavy metals to create highly multiplexed, parallel immunoassays.

Mullis was a member of the [[USA Science and Engineering Festival]]'s Advisory Board.<ref>{{Cite web |url=http://www.usasciencefestival.org/about/advisors |title=Advisors |publisher=USA science festival |access-date=July 27, 2010 |url-status=dead |archive-url=https://web.archive.org/web/20100421005310/http://www.usasciencefestival.org/about/advisors/ |archive-date=April 21, 2010 |df=mdy-all}}</ref> In 2014, he was named a distinguished researcher at the [[Children's Hospital Oakland Research Institute]] in [[Oakland, California]].<ref name="Biography">{{cite web |url=http://www.karymullis.com/biography.shtml |title=Biography |website=Karymullis.com |access-date=May 9, 2016}}</ref>

=== PCR (polymerase chain reaction) and other inventions ===

{{Main|Taq polymerase|History of polymerase chain reaction}}
In 1983, Mullis was working for Cetus Corporation as a chemist.<ref name="Yoffe 1994" /> Mullis recalled that, while driving in the vicinity of his country home in [[Mendocino County]] (with his girlfriend, who also was a chemist at Cetus), 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 that would allow rapid amplification of a small stretch of DNA and become a standard procedure in molecular biology laboratories.<ref name="Yoffe 1994" /> Longtime professional benefactor and supervisor Thomas White reassigned Mullis from his usual projects to concentrate on PCR full-time after the technique was met with skepticism by their colleagues.{{r|Yoffe 1994|McDonald}} Mullis succeeded in demonstrating PCR on December 16, 1983, but the staff remained circumspect as he continued to produce ambiguous results amid alleged methodological problems, including a perceived lack of "appropriate controls and repetition."{{r|Yoffe 1994|McDonald}} In his Nobel Prize lecture, he remarked that the December 16 breakthrough did not make up for his girlfriend breaking up with him: "I was sagging as I walked out to my little silver [[Honda Civic]]. Neither [assistant] Fred, empty [[Beck's Brewery|Beck's]] bottles, nor the sweet smell of the dawn of the age of PCR could replace Jenny. I was lonesome."<ref name="Yoffe 1994" />

Other Cetus scientists who were regarded as "top-notch experimentalists",<ref name="McDonald"/> including Randall Saiki, Henry Erlich, and [[Norman Arnheim]], were placed on parallel PCR projects to work on determining if PCR could amplify a specific human gene (betaglobin) from genomic DNA. Saiki generated the needed data and Erlich authored the first paper to include use of the technique,<ref name="Wade 1998" /> while Mullis was still working on the paper that would describe PCR itself.<ref name="Yoffe 1994" /> Mullis's 1985 paper with Saiki and Erlich, "Enzymatic Amplification of β-globin Genomic Sequences and Restriction Site Analysis for Diagnosis of Sickle Cell Anemia" — the polymerase chain reaction invention (PCR) — was honored by a Citation for Chemical Breakthrough Award from the Division of History of Chemistry of the American Chemical Society in 2017.<ref name="breakthrough">{{Cite web |title=Citations for Chemical Breakthrough Awards 2017 Awardees |url=http://www.scs.illinois.edu/~mainzv/HIST/awards/CCB-2017_Awardees.php |publisher=Division of History of Chemistry of the American Chemical Society |access-date=12 March 2018}}</ref><ref>{{Cite journal |last1=Saiki |first1=R. |last2=Scharf |first2=S. |last3=Faloona |first3=F. |last4=Mullis |first4=K. |last5=Horn |first5=G. |last6=Erlich |first6=H. |last7=Arnheim |first7=N |title=Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia |journal=Science |date=20 December 1985 |volume=230 |issue=4732 |pages=1350–1354 |doi=10.1126/science.2999980 |pmid=2999980 |bibcode=1985Sci...230.1350S}}</ref>

A drawback of the technique was that the DNA polymerase in the reaction was destroyed by the high heat used at the start of each replication cycle and had to be replaced. In 1986, Saiki started to use ''[[Thermophilus aquaticus]]'' (Taq) [[DNA polymerase]] to amplify segments of DNA. The Taq polymerase was heat resistant and needed to be added to the reaction only once, making the technique dramatically more affordable and subject to automation. This modification of Mullis's invention revolutionized  biochemistry, molecular biology, [[genetics]], medicine, and [[forensics]]. UC Berkeley biologist David Bilder said, "PCR revolutionized everything. It really superpowered molecular biology—which then transformed other fields, even distant ones like ecology and [[evolution]]. … It’s impossible to overstate PCR’s impact. The ability to generate as much DNA of a specific sequence as you want, starting from a few simple chemicals and some temperature changes—it’s just magical."<ref name="McDonald"/> Although he received a $10,000 bonus from Cetus for the invention, the company's later sale of the patent to [[Roche Molecular Systems]] for $300&nbsp;million would lead Mullis to condemn White and members of the parallel team as "vultures."{{r|Yoffe 1994|McDonald}}

Mullis also invented a UV-sensitive plastic that changes color in response to light.<ref name=SalemBioEncyc>{{cite book |author=Vigue, C.L. |title=Salem Press Biographical Encyclopedia: Kary B. Mullis |publisher=Salem Press, a division of EBSCO |date=2021}}</ref>

He founded Altermune LLC in 2011 to pursue new ideas on the immune system.<ref name="Business Wire">{{cite press release |author1=<!--Staff writer; no byline--> |title=Loxbridge and Dr Kary Mullis Announce the Formation of Altermune Technologies with $7m Seed Investment |url=https://www.businesswire.com/news/home/20110922005770/en/Loxbridge-and-Dr-Kary-Mullis-Announce-the-Formation-of-Altermune-Technologies-with-7m-Seed-Investment |website=Business Wire |date=22 September 2011}}</ref> Mullis described the company's product thusly:
{{Blockquote |style=font-size:100% |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.<ref name="Nobel bio" />}}

In a [[TED Talk]], Mullis describes how the US government paid $500,000 for Mullis to use this new technology against [[anthrax]]. He said the treatment was 100% effective, compared to the previous anthrax treatment which was 40% effective.<ref name="Mullis Feb 2009">{{Cite AV media |people=Mullis, Kary |url=https://www.ted.com/talks/kary_mullis_a_next_gen_cure_for_killer_infections/transcript |title=A next-gen cure for killer infections |type=video with transcript |website=TED.com |date=February 2009 |access-date=July 27, 2010}}</ref>

Another proof-of-principle of this technology, re-targeting pre-existing antibodies to the surface of a pathogenic strep bacterium using an alpha-gal modified aptamer ("alphamer"), was published in 2015 in collaboration with scientists at the University of California, San Diego.<ref>{{Cite press release |first=Heather |last=Buschman |title=Molecular Homing Beacon Redirects Human Antibodies to Fight Pathogenic Bacteria |url=http://ucsdnews.ucsd.edu/pressrelease/molecular_homing_beacon_redirects_human_antibodies_to_fight_pathogenic_bact |publisher=[[University of California, San Diego]] |date=May 6, 2015 |access-date=July 6, 2017}}</ref><ref>{{Cite journal |pmid=25940316 |title=Retargeting pre-existing human antibodies to a bacterial pathogen with an alpha-Gal conjugated aptamer |doi=10.1007/s00109-015-1280-4 |pmc=4469262 |volume=93 |issue=6 |year=2015 |journal=J. Mol. Med. |pages=619–31 |last1=Kristian |first1=SA |last2=Hwang |first2=JH |last3=Hall |first3=B |last4=Leire |first4=E |last5=Iacomini |first5=J |last6=Old |first6=R |last7=Galili |first7=U |last8=Roberts |first8=C |last9=Mullis |first9=KB |last10=Westby |first10=M |last11=Nizet |first11=V}}</ref> Mullis said he was inspired to fight this particular strep bacterium because it had killed his friend.<ref name="Mullis Feb 2009" />

=== Accreditation of the PCR technique ===

{{See also|History of polymerase chain reaction}}

A concept similar to that of PCR had been described before Mullis's work. Nobel laureate [[H. Gobind Khorana]] and [[Kjell Kleppe]], a Norwegian scientist, authored a paper 17 years earlier describing a process they termed "repair replication" in the ''[[Journal of Molecular Biology]]''.<ref name=":0">{{Cite journal |last1=Kleppe |first1=K. |last2=Ohtsuka |first2=E. |last3=Kleppe |first3=R. |last4=Molineux |first4=I. |last5=Khorana |first5=H. G. |author-link5=Har Gobind Khorana |year=1971 |title=Studies on polynucleotides *1, *2XCVI. Repair replication of short synthetic DNA's as catalyzed by DNA polymerases |journal=Journal of Molecular Biology |volume=56 |issue=2 |pages=341–361 |doi=10.1016/0022-2836(71)90469-4 |pmid=4927950}}</ref> Using repair replication, Kleppe duplicated and then quadrupled a small synthetic molecule with the help of two primers and DNA polymerase. The method developed by Mullis used repeated thermal cycling, which allowed the rapid and exponential amplification of large quantities of any desired DNA sequence from an extremely complex template. Later a heat-stable DNA polymerase was incorporated into the process.

His co-workers at Cetus contested the notion that Mullis was solely responsible for the idea of using [[Taq polymerase]] in PCR.{{Citation needed|date=December 2022}} However, biochemist [[Richard T. Pon]] has written that the "full potential [of PCR] was not realized" until Mullis's work in 1983,<ref name="Pon 2002">{{Cite book |last=Pon |first=Richard T. |editor-last1=Khudyakov |editor-first1=Yury E. |editor-last2=Fields |editor-first2=Howard A. |title=Artificial DNA: Methods and Applications |page=20 |isbn=978-1-4200-4016-6 |year=2002 |publisher=[[CRC Press]] |location=Boca Raton, Fla. |chapter=Chemical Synthesis of Oligonucleotides: From Dream to Automation}}</ref> and journalist [[Michael Gross (science writer)|Michael Gross]] states that Mullis's colleagues failed to see the potential of the technique when he presented it to them.<ref name="Gross 2001">{{Cite book |title=Life on the Edge: Amazing Creatures Thriving in Extreme Environments |year=2001 |isbn=978-0-7382-0445-1 |last1=Gross |first1=Michael |publisher=Basic Books |location=New York |page=103 |url=https://archive.org/details/lifeonedgeamazin0000gros_t5j2/page/103/mode/1up?view=theater |url-access=registration}}</ref>{{Synthesis inline|date=December 2022}} As a result, some controversy surrounds the balance of credit that should be given to Mullis versus the team at Cetus.<ref name="Wade 1998" /> In practice, credit has accrued to both the inventor and the company (although not its individual workers) in the form of a Nobel Prize and a $10,000 Cetus bonus for Mullis and $300&nbsp;million for Cetus when the company sold the patent to Roche Molecular Systems. After [[DuPont (1802–2017)|DuPont]] lost out to Roche on that sale, the company unsuccessfully disputed Mullis's patent on the alleged grounds that PCR had been previously described in 1971.<ref name="Yoffe 1994" /> Mullis and Erlich took Cetus' side in the case, and Khorana refused to testify for DuPont; the jury upheld Mullis's patent in 1991.<ref name="Yoffe 1994" /> However, in February 1999, the patent of Hoffman-La Roche (United States Patent No. 4,889,818) was found by the courts to be unenforceable, after Dr. Thomas Kunkel testified in the case ''Hoffman-La Roche v. Promega Corporation''<ref name=Roche_v_Promega>{{Cite web |url=https://www.casemine.com/judgement/us/5914b97badd7b0493478b532 |title=Hoffmann-La Roche, Inc. v. Promega Corporation, (N.D.Cal. 2001) |publisher=Casemine |author=United States District Court, N.D. California. |date=July 26, 2001 |access-date=April 8, 2018}}</ref> on behalf of the defendants ([[Promega Corporation]]) that "prior art" (i.e. articles on the subject of Taq polymerase published by other groups prior to the work of Gelfand and Stoffel, and their patent application covering the purification of Taq polymerase) existed, in the form of two articles, published by Alice Chien et al. in 1976,<ref name="Chien">{{Cite journal |last1=Chien |first1=A. |last2=Edgar |first2=D.B. |last3=Trela |first3=J.M. |title=Deoxyribonucleic acid polymerase from the extreme thermophile Thermus aquaticus |journal=J. Bacteriol. |volume=127 |issue=3 |pages=1550–1557 |year=1976 |pmid=8432 |pmc=232952 |doi=10.1128/jb.127.3.1550-1557.1976}}</ref> and A. S. Kaledin et al. in 1980.<ref name="Kaledin">{{Cite journal |last1=Kaledin |first1=A.S. |last2=Sliusarenko |first2=A.G. |last3=Gorodetskii |first3=S.I. |title=Isolation and properties of DNA polymerase from extreme thermophylic bacteria Thermus aquaticus YT-1 |journal=[[Biokhimiia]] |volume=45 |issue=4 |pages=644–651 |year=1980 |pmid=7378495}}</ref>

The anthropologist [[Paul Rabinow]] wrote a book on the history of the PCR method in 1996,<ref name="Bilsker 1998">{{Cite journal |first=Richard |last=Bilsker |url=http://www.hyle.org/journal/issues/4/bilsker.htm |title=Ethnography of a Nobel Prize |journal=Hyle: International Journal for Philosophy of Chemistry |volume=4 |issue=2 |year=1998 |pages=167–169 |issn=1433-5158 |access-date=July 27, 2010}}</ref> in which he discusses whether Mullis "invented" PCR or merely came up with the concept of it.<ref name="Rabinow p4">{{cite book |last=Rabinow |first=Paul |title=Making PCR: A Story of Biotechnology |publisher=University of Chicago Press |year=1996 |pages=4–5 |isbn=0-226-70147-6 |url=https://archive.org/details/makingpcrstoryof00rabi/page/4/mode/1up?view=theater |url-access=registration}}</ref>{{Explain|date=December 2022}}