Overexpression of human testis antigens in Escherichia coli host cells is influenced by site of expression and the induction temperature.
A panel of twenty human testis cDNA clones were expressed in an Escherichia coli expression system and six clones were found to express identifiable fusion polypeptides. Expression was found to be influenced not only by the site of localization of the polypeptide in the host cells, but also by the temperature used for induction. This emphasized the need for cytoplasmic and periplasmic expression of new antigens of unknown properties, as well as the use of temperatures of 30 degrees C or lower. A majority of the expressed polypeptides were mainly in an insoluble form. By reducing the induction temperature to 30 degrees C production of the soluble fraction was further improved.
Mburu
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DN
Department of Biological Sciences, University of Newcastle, Callaghan, Australia. david.mburu@mcmail.vanderbilt.edu
Roberts
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TK
Boettcher
B
B
D000941:Antigens / Q000235:genetics*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000235:genetics*; D005786:Gene Expression Regulation; D006801:Humans; D008297:Male; D009842:Oligopeptides; D010455:Peptides / Q000235:genetics / Q000276:immunology; D013696:Temperature; D013737:Testis / Q000276:immunology*
D000941:Antigens; D018076:DNA, Complementary; D009842:Oligopeptides; D010455:Peptides; C085706:FLAG peptide