Douglas Prasher

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Douglas C. Prasher (born August 1951) is an American molecular biologist. He is known for his work to clone and sequence the genes for the photoprotein aequorin [1] and green fluorescent protein (GFP) [2] and for his proposal to use GFP as a tracer molecule.[3]

Career[edit]

Prasher received his Ph.D. in Biochemistry from the Ohio State University in 1979. From 1979 to 1983, he worked in genetics and biochemistry research at the University of Georgia, where he identified the gene sequence for aequorin [1] .[4] He then joined the Biology Department of the Marine Biological Laboratory, Woods Hole, Massachusetts where he studied bioluminescence. In 1988, he received a two-year, $200,000 grant from the American Cancer Society to clone the gene for green fluorescent protein (GFP), the protein that gives the jellyfish its glow. Prasher succeeded in this project, and later shared his findings with Martin Chalfie and Roger Y. Tsien after each scientist had communicated with him.[5][6] In subsequent years, Prasher provided the clone to hundreds of scientists.[citation needed]

Reports that Prasher had difficulty in achieving fluorescence of GFP in other species in recombinant studies [7] are inaccurate, as Prasher had successfully worked with the Chalfie group to show recombinant expression in the bacterium E. coli and the nematode C. elegans,[8] and later in the plant Arabidopsis thaliana.[9] By the time Prasher's ACS funding ended, he had isolated a partial, but almost complete gfp cDNA, with 965 bases out of the 10,50 bases of the corresponding mRNA.[2] It would require construction of another cDNA library during the following (non-funded) year for Prasher to isolate a full-length cDNA clone, although it must be noted that this partial cDNA clone was subsequently used and found to be sufficient for successful heterologous expression in E. coli, C. elegans [8] and A. thaliana.[9] By this time Prasher could not afford to devote limited resources to expression studies in E. coli.[citation needed] It wasn't until the Nobel Prize announcement that it became clear how unfortunate this had been. Chalfie and Tsien went on to their successful expression studies. GFP has subsequently found application as a biochemical tracer such as in fluorescent studies of gene expression.

Prasher had applied to the National Institutes of Health for funding but had been turned down, and by the time he was undergoing tenure review, he had decided to leave academia.[citation needed] Subsequently, Prasher worked for the Animal & Plant Health Inspection Service,[1] U.S. Department of Agriculture at its Otis Plant Protection Center in Cape Cod, Massachusetts as a population geneticist, and was later transferred to the Plant Germplasm Quarantine & Biotechnology Laboratory in Beltsville, Maryland. After working conditions deteriorated at the Beltsville location, he went to work for NASA subcontractor AZ Technology in Huntsville, Alabama working on an existing project to develop lab-on-a-chip devices to monitor cabin environment and to perform human diagnostics during long-term spaceflight.[citation needed] However, he lost his job after 1.5 years when NASA reorganized and canceled the project.[10][11]

On 8 October 2008, the 2008 Nobel Prize in Chemistry was awarded to Osamu Shimomura, Chalfie, and Tsien for their work on GFP.[12] Prasher was not included among the Nobel laureates, as only 3 individuals can share in a single Nobel Prize. Chalfie said of Prasher's contribution:

"(Douglas Prasher's) work was critical and essential for the work we did in our lab. They could've easily given the prize to Douglas and the other two and left me out."[13]

Tsien also agreed that they couldn't have done it without Prasher and "Doug Prasher had a very important role."[14]

In an October 9, 2008 phone interview with National Public Radio (NPR) and October 14, 2008 TV interview with Inside Edition, Prasher reported that he was unable to find a job in science, his life savings had run out and that he was working as a courtesy shuttle bus driver for a Toyota dealership in Huntsville, Alabama at $8.50 an hour.[6][11][13][14][15] In the NPR broadcast, one of his former colleagues called Prasher's current situation a "staggering waste of talent."[6] Prasher has stated his wish to resume a career in science, but not particularly with jellyfish.[10] He has publicly expressed his pleasure at learning of the awarding of the Nobel Prize to Shimomura, Chalfie and Tsien:

"I'm really happy for them. I was really surprised that particular topic carried that much weight."[10]

Chalfie and Tsien invited Prasher and his wife, Virginia Eckenrode, to attend the Nobel Prize ceremony, as their guests and at their expense.[16] All three of the 2008 Chemistry laureates thanked Prasher in their speeches.[17]

In June 2010, Prasher was finally able to return to science, working for Streamline Automation in Huntsville until December 2011, then from 2012 on, in Roger Tsien's lab at the University of California in San Diego[18]

Publications[edit]

  • Prasher, D., McCann, R.O., Cormier, M.J., Cloning and expression of the cDNA coding for aequorin, a bioluminescent calcium-binding protein. Biochem. Biophys. Res. Comm., 126, 1259-1268 (1985).
  • Richard, J.P., Prasher, D.C., Ives, D.H., Frey, P.A., Chiral [18O]phosphorothioates. The stereochemical course of thiophosphoryl group transfer catalyzed by nucleoside phosphotransferase. J. Biol. Chem., 254(11), 4339-4341 (1979).
  • Prasher, D.C., Carr, M.C., Ives, D.H., Tsai, T.C., Frey, P.A., Nucleoside phosphotransferase from barley. Characterization and evidence for ping pong kinetics involving phosphoryl enzyme. J. Biol. Chem., 257(9), 4931-4939 (1982).
  • Prasher, D.C., Conarro, L., Kushner, S.R., Amplification and purification of exonuclease I from Escherichia coli K12. J. Biol. Chem., 258(10), 6340-6343 (1983)
  • Prasher, D.C., McCann, R.O., Longiaru, M., Cormier, M.J., Sequence comparisons of complementary DNAs encoding aequorin isotypes. Biochemistry, 26(5), 1326-1332 (1987).
  • Phillips, G.J., Prasher, D.C., Kushner, S.R., Physical and biochemical characterization of cloned sbcB and xonA mutations from Escherichia coli K-12. J. Bacteriol., 170(5), 2089-2094 (1988).
  • Cormier, M.J., Prasher, D.C., Longiaru, M., McCann, R.O., The enzymology and molecular biology of the Ca2+-activated photoprotein, aequorin. Photochem. Photobiol., 49(4), 509-512 (1989).
  • Prasher, D.C., O'Kane, D., Lee, J., Woodward, B., The lumazine protein gene in Photobacterium phosphoreum is linked to the lux operon. Nucleic Acids Res., 18(21), 6450 (1990).
  • O'Kane, D.J., Woodward, B., Lee, J., Prasher, D.C., Borrowed proteins in bacterial bioluminescence. Proc. Natl. Acad. Sci. USA, 88(4), 1100-1104 (1991).
  • O'Kane, D.J., Prasher, D.C., Evolutionary origins of bacterial bioluminescence. Mol. Microbiol., 6(4), 443-449 (1992).
  • Prasher, D.C., Eckenrode, V.K., Ward, W.W., Prendergast F.G., Cormier, M.J., Primary structure of the Aequorea victoria green-fluorescent protein. Gene, 111(2), 229-233 (1992).
  • Hannick, L.I., Prasher, D.C., Schultz, L.W., Deschamps, J.R., Ward, K.B., Preparation and initial characterization of crystals of the photoprotein aequorin from Aequorea victoria. Proteins, 15(1), 103-107 (1993).
  • Cody, C.W., Prasher, D.C., Westler, W.M., Prendergast, F.G., Ward, W.W., Chemical structure of the hexapeptide chromophore of the Aequorea green-fluorescent protein. Biochemistry, 32(5), 1212-1218 (1993).
  • Chalfie, M., Tu, Y., Euskirchen, G., Ward, W.W., Prasher, D.C., Green fluorescent protein as a marker for gene expression. Science, 263(5148), 802-805 (1994).
  • Heim, R., Prasher, D.C., Tsien, R.Y., Wavelength mutations and posttranslational autoxidation of green fluorescent protein. Proc. Natl. Acad. Sci. USA, 91(26), 12501-12504 (1994).
  • Prasher, D.C., Using GFP to see the light. Trends Genet., 11(8), 320-323 (1995).
  • Haseloff, J., Siemering, K.R., Prasher, D.C., Hodge, S., Removal of a cryptic intron and subcellular localization of green fluorescent protein are required to mark transgenic Arabidopsis plants brightly. Proc. Natl. Acad. Sci. USA, 94(6), 2122-2127 (1997).
  • Bernon, G., Schander, C., Prasher, D., Robinson, D., Survey and status of terrestrial slugs in North America American Malacological Society Abstracts 2000, 41 (2000).
  • Barr, NB, Cook, A., Elder, P., Molongoski, J., Prasher, D., Robinson D.G. Application of a DNA barcode using the 16S rRNA gene to diagnose pest Arion species in the USA. " J. Moll. Stud. 75: 187-191 (2009).

References[edit]

  1. ^ a b Prasher, D C; R O McCann; M Longiaru; M J Cormier (1987-03-10). "Sequence comparisons of complementary DNAs encoding aequorin isotypes". Biochemistry 26 (5): 1326–1332. ISSN 0006-2960. PMID 2882777. 
  2. ^ a b Prasher, D C; V K Eckenrode; W W Ward; F G Prendergast; M J Cormier (1992-02-15). "Primary structure of the Aequorea victoria green-fluorescent protein". Gene 111 (2): 229–233. doi:10.1016/0378-1119(92)90691-H. ISSN 0378-1119. PMID 1347277. 
  3. ^ Prasher, D C (1995-08). "Using GFP to see the light". Trends in genetics: TIG 11 (8): 320–323. ISSN 0168-9525. PMID 8585130. 
  4. ^ Matthew Herper (2008-10-08). "Biotech's Glowing Breakthrough Wins Nobel Prize". Forbes. Retrieved 2008-10-13. 
  5. ^ Kenneth Chang (2008-10-09). "Three Chemists Win Nobel Prize". New York Times. Retrieved 2008-10-11. 
  6. ^ a b c Dan Charles (2008-10-09). "Glowing Gene's Discoverer Left Out Of Nobel Prize". Morning Edition (National Public Radio [NPR]). Retrieved 2008-10-05. 
  7. ^ Tina Hesman Saey (2008-10-25). "Nobel Prize in Chemistry Commends Finding and Use of Green Fluorescent Protein". Science News. Retrieved 2008-10-11. 
  8. ^ a b Chalfie, M; Y Tu; G Euskirchen; W W Ward; D C Prasher (1994-02-11). "Green fluorescent protein as a marker for gene expression". Science 263 (5148): 802–805. doi:10.1126/science.8303295. ISSN 0036-8075. PMID 8303295. 
  9. ^ a b Haseloff, J; K R Siemering; D C Prasher; S Hodge (1997-03-18). "Removal of a cryptic intron and subcellular localization of green fluorescent protein are required to mark transgenic Arabidopsis plants brightly". Proceedings of the National Academy of Sciences of the United States of America 94 (6): 2122–2127. ISSN 0027-8424. PMC 20051. PMID 9122158. 
  10. ^ a b c Steve Doyle (2008-10-10). "Local biochemist had hand in Nobel". Huntsville Times. Retrieved 2008-10-11. 
  11. ^ a b Associated Press (2008-10-11). "Van driver's work in Mass. aided Nobel winners". Boston Globe. Retrieved 2014-03-17. 
  12. ^ "The Nobel Prize in Chemistry 2008" (Press release). The Royal Swedish Academy of Sciences. 8 October 2008. Retrieved 2008-10-11. 
  13. ^ a b Aaron Gouveia (2008-10-11). "Shuttle driver reflects on Nobel snub". Cape Cod Times. Retrieved 2008-10-11. 
  14. ^ a b Jim Moret (2008-10-14). "Genius Behind the Wheel - Why is a genius scientist driving a bus?". Inside Edition. Retrieved 2008-10-16. 
  15. ^ Philip Sherwell (2008-10-11). "The scientist, the jellyfish protein and the Nobel prize that got away". The Daily Telegraph. Retrieved 2008-10-11. 
  16. ^ Steve Doyle (2008-12-04). "Van driver to attend Nobel ceremony". Huntsville Times. Retrieved 2008-12-11. 
  17. ^ Lee Roop (2008-12-18). "'Magical' Nobel trip could lead to new opportunities". Huntsville Times. Retrieved 2009-02-04. 
  18. ^ Douglas Prasher profile

External links[edit]