SELECTION OF RECOMBINANT VACCINIA VIRUSES (TIAN TAN STRAIN) EXPRESSING HEPATITIS B VIRUS SURFACE ANTIGEN BY USING β-GALACTOSIDASE AS A MARKER*  被引量:1

SELECTION OF RECOMBINANT VACCINIA VIRUSES (TIAN TAN STRAIN) EXPRESSING HEPATITIS B VIRUS SURFACE ANTIGEN BY USING β-GALACTOSIDASE AS A MARKER*

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作  者:刘庚起 曹旭 朱既明 任贵方 李红梅 解燕乡 彭薇 

机构地区:[1]Institute of Virology, Chinese Academy of Preventive Medicine, Beijing 100052, PRC

出  处:《Science China Chemistry》1990年第2期188-197,共10页中国科学(化学英文版)

基  金:Project supported by the National Natural Science Foundation of China.

摘  要:We constructed a plasmid that contains a small piece of DNA with two vaccinis promoters running in opposite directions——a promoter from a late gene encoding an 11 K polypeptide (P11) and a promoter from an early gene encoding 25K(P25). These promoters were isolated from the Tian Tan strain of vaccinia virus and were flanked by the thymidine kinase (TK) sequence of the same virus. Genes encoding the hepatitis B virus surface antigen (HBsAg) and the Rscherichia coli β-galactosidase (LacZ) were inserted downstream of the 11 K and 25 K promoters respectively so that coexpression plasmids were constructed. Recombinant vaccinis Viruses were selected directly by picking blue plaques formed under overlaying agarose medium containing X-gal. HBsAg was expressed to high level by these recombinant viruses. These recombinant viruses showed reduced virulence on rabbit skin and induced anti-HBs after intrsdermal inoculation of rabbits.We constructed a plasmid that contains a small piece of DNA with two vaccinis promoters running in opposite directions——a promoter from a late gene encoding an 11 K polypeptide (P11) and a promoter from an early gene encoding 25K(P25). These promoters were isolated from the Tian Tan strain of vaccinia virus and were flanked by the thymidine kinase (TK) sequence of the same virus. Genes encoding the hepatitis B virus surface antigen (HBsAg) and the Rscherichia coli β-galactosidase (LacZ) were inserted downstream of the 11 K and 25 K promoters respectively so that coexpression plasmids were constructed. Recombinant vaccinis Viruses were selected directly by picking blue plaques formed under overlaying agarose medium containing X-gal. HBsAg was expressed to high level by these recombinant viruses. These recombinant viruses showed reduced virulence on rabbit skin and induced anti-HBs after intrsdermal inoculation of rabbits.

关 键 词:RECOMBINANT VACCINIA viruses HEPATITIS B Β-GALACTOSIDASE 

分 类 号:O6[理学—化学]

 

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