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Nucleic Acids Research, 1983, Vol. 11, No. 11 3581-3591
© 1983

MOLECULAR BIOLOGY

Direct expression of hepatitis B surface antigen gene in E coli

Yukio Fujisawa, Yasuaki Ito, Reiko Sasada, Yoshitaka Ono, Koichi Igarashi, Ryuji Marumoto, Masakazu Kikuchi and Yukio Sugino

Biotechnology Laboratories, Central Research Division, Takeda Chemical Industries Ltd. Osaka 532, Japan

Received March 21, 1983. Accepted May 6, 1983.

A 809 bp Sau 3A – Hpa I fragment containing a complete HBsAg gene and fragments 744 bp Hinc II – Hpa I and 712 bp Xba I – Hpa I containing a truncated HBsAg gene lacking the sequence encoding the NH2terminal hydrophobic domain were prepared from a composite plasmid pHBV933 containing the 3.2 kb Eco RI DNA fragment of the entire HBVminusbadw genome and inserted into an expression vector pTRP801 to give plasmids pTRP SS-6, pTRP SS-39, and pTRP SS–50, respectively. The growth of a recombinant having pTRP SS-6 was greatly inhibited and the transformant expressed a low level of HBsAg, which is reactive to human anti-HBsAg antibody. Interestingly, the growth of transformants harbouring pTRP SS-39 and pTRP SS–50 was not inhibited and these transformants expressed a considerable level of the HBsAg. Minicells harbouring pTRP SS–6, pTRP SS–39, and pTRP SS–50 formed specific polypeptides of about 24 K, 23 K, and 22 K daltons, respectively.


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