Increased expression of hepatitis B virus surface antigen gene in Escherichia coli by IS1 insertion |
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Authors: | Kazuo Nakahama Yasuaki Itoh Yukio Fujisawa Masakazu Kikuchi |
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Affiliation: | (1) Biotechnology Laboratories, Central Research Division, Takeda Chemical Industries Ltd., 532 Osaka, Japan;(2) Biotechnology laboratories, Central Research Division, Takeda Chemical Industries Ltd., 17-85, Jusohonmachi 2-chome, Yodogawa-ku, 532 Osaka, Japan |
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Abstract: | Summary We selected faster growing colonies of Escherichia coli harbouring an expression plasmid for hepatitis B virus surface antigen (HBsAg) gene after mutagenesis. Among these colonies, three were found to produce an increased level of HBsAg as a consequence of alteration of the plasmid. Analysis of this plasmid showed that an insertion sequence, IS1, was inserted into a middle region of the HBsAg gene (codon for Pro 127) to generate a termination codon 20 bp downstream from the junction site between the HBsAg gene and the left end of IS1. Insertion of a chemically synthesized termination codon into the same region of the HBsAg gene also increased the expression of the HBsAg gene. These results suggest that HBsAg lacking the COOH-terminal region is produced at a high level because it does not inhibit the growth of the host. |
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