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Choosing Between Yeast and Bacterial Expression Systems: Yield DependentGreen 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.
Document ID
20020051097
Acquisition Source
Marshall Space Flight Center
Document Type
Preprint (Draft being sent to journal)
Authors
Miller, Rebecca S.
(Alabama Univ. Huntsville, AL United States)
Malone, Christine C.
(Universities Space Research Association Huntsville, AL United States)
Moore, Blake P.
(Alabama Univ. Huntsville, AL United States)
Burk, Melissa
(Morgan Research Corp. Huntsville, AL United States)
Crawford, Lisa
(Universities Space Research Association Huntsville, AL United States)
Karr, Laurel J.
(NASA Marshall Space Flight Center Huntsville, AL United States)
Curreri, Peter A.
Date Acquired
August 20, 2013
Publication Date
January 1, 2002
Subject Category
Life Sciences (General)
Meeting Information
Meeting: Current Topics in Gene Expression Systems Meeting
Location: San Diego, CA
Country: United States
Start Date: March 25, 2002
Distribution Limits
Public
Copyright
Work of the US Gov. Public Use Permitted.

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