Cloning of two protease genes fromRhodocyclus gelatinosa APR 3-2 and their expression inEscherichia coli |
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Authors: | Dr Kazutoshi Ito Shin-Ichi Sakakibara Kazuo Izaki |
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Institution: | (1) Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku University, 980 Sendai, Japan |
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Abstract: | Two different protease genes were cloned fromRhodocyclus gelatinosa APR 3-2 inEscherichia coli HB 101/ with pBR329 or its derivatives. The recombinant plasmids designated as pRP100 and pRP300 contained 11.2 and 10.6 kb DNA fragments, respectively. The differences of both plasmids in restriction enzyme maps indicate that these plasmids contained different protease genes. DNA fragments coding for protease, 6.4 kb and 4.5 kb from pRP100 and pRP300, were subcloned into pRP329 and designated as pRP101 and pRP301, respectively. The two cloned proteases were excreted in culture medium ofE. coli, and ß-lactamase ofE. coli, which was originally localized in periplasmic space, was also excreted in the medium. |
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