+ Visit NASA.gov + Contact NASA   + Home Collection > NASA  Author > Burk, Melissa  Author > Crawford, Lisa  Author > Karr, Laurel J.  Author > Malone, Christine C.  Author > Miller, Rebecca S.  Author > Moore, Blake P.  NASA Center > Marshall Space Flight Center  Publication Year > 2001-2010 > 2002  Subject > J-L > Life Sciences (general)  Availability Options > Order From Other Sources  Item/Media Type > Preprint  Note: Start a new navigation search by selecting a link above     Previous Record | Next Record + Back to Results   + Printer Friendly Title: Choosing Between Yeast and Bacterial Expression Systems: Yield Dependent Author(s): Miller, Rebecca S.; Malone, Christine C.; Moore, Blake P.; Burk, Melissa; Crawford, Lisa; Karr, Laurel J. Abstract: Green fluorescent protein (GFP) is a naturally occurring fluorescent protein isolated from the jellyfish Aequorea victoria. The intrinsic fluorescence of the protein is due to a chromophore located in the center of the molecule. Its usefulness has been established as a marker for gene expression and localization of gene products. GFP has recently been utilized as a model protein for crystallization studies at NASA/MSFC, both in earth-based and in microgravity experiments. Because large quantities of purified protein were needed, the cDNA of GFP was cloned into the Pichia pastoris pPICZ(alpha) C strain, with very little protein secreted into the media. Microscopic analysis prior to harvest showed gigantic green fluorescent yeast, but upon harvesting most protein was degraded. Trial fermentations of GFP cloned into pPICZ A for intracellular expression provided unsatisfactory yield. GFP cloned into E, coli was overexpressed at greater than 150 mg/liter, with purification yields at greater than 100mg/liter. NASA Center: Marshall Space Flight Center Publication Date: [2002] Document Source: Other Sources No Digital Version Available: Go to Tips On Ordering Available Data: Abstract Only Document ID: 20020051097 Publication Information: Number of Pages = 1 Meeting Information: Current Topics in Gene Expression Systems Meeting, FROM, 25 Mar. 2002, San Diego, CA, United States Keywords: PROTEIN CRYSTAL GROWTH; YEAST; BACTERIA; FLUORESCENCE; GRAVITATIONAL EFFECTS; GENE EXPRESSION; PURIFICATION; MICROGRAVITY; FERMENTATION; ESCHERICHIA; CHROMOPHORES; Accessibility: Unclassified; No Copyright; Unlimited; Publicly available; Updated/Added to NTRS: 2004-11-03 + Back to Top + Sponsored by the NASA Scientific and Technical    Information Program + 2004 Vision for Space Exploration + Freedom of Information Act + NASA Web Privacy Policy and Important Notices + NASA Disclaimers, Copyright Notice,    and Terms and Conditions of Use NASA Official: Calvin Mackey Page Curator: NASA Center for AeroSpace Information (help@sti.nasa.gov) Last Updated: July 5, 2007