Fujisawa Y, Ito Y, Sasada R, Ono Y, Igarashi K, Marumoto R, Kikuchi M, Sugino Y
Nucleic Acids Res. 1983 Jun 11;11(11):3581-91. doi: 10.1093/nar/11.11.3581.
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 NH2-terminal hydrophobic domain were prepared from a composite plasmid pHBV933 containing the 3.2 kb Eco RI DNA fragment of the entire HBV/adw 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.
从含有整个乙肝病毒/adw基因组3.2 kb Eco RI DNA片段的复合质粒pHBV933中制备出一个包含完整乙肝表面抗原（HBsAg）基因的809 bp Sau 3A - Hpa I片段，以及两个分别包含截短型HBsAg基因（缺少编码NH2末端疏水结构域序列）的片段，即744 bp Hinc II - Hpa I片段和712 bp Xba I - Hpa I片段，并将它们分别插入表达载体pTRP801，得到质粒pTRP SS - 6、pTRP SS - 39和pTRP SS - 50。携带pTRP SS - 6的重组体生长受到极大抑制，转化体表达的HBsAg水平较低，该HBsAg与人抗HBsAg抗体有反应。有趣的是，携带pTRP SS - 39和pTRP SS - 50的转化体生长未受抑制，且这些转化体表达了相当水平的HBsAg。携带pTRP SS - 6、pTRP SS - 39和pTRP SS - 50的微小细胞分别形成了约24 kDa、23 kDa和22 kDa的特异性多肽。
