[Rapid construction of recombinant baculovirus carrying hepatitis B virus surface antigen gene by using a novel baculovirus shuttle vector(Bacmid)].
In order to express the HBV surface antigen (HBV) efficiently by using the Bac-to-Bac system and to produce HBsAg for diagnosis and vaccine development of HBV, a recombinant donor plasmid pFB-BS carrying HBsAg gene was constructed. The gene was inserted into shuttle plasmid (Bacmid) in E. coli DH10Bac with the help of Tn7 transposition system. The recombinant Bacmid was examined by PCR and transfected to Sf9 cells. HBsAg was expressed under the control of polyhedrin promoter of baculovirus in insect cells and was identified by ELISA. The quantity of the expression reached to 2 micrograms/10(6) cells and most production was within cells. Our data suggested that the Bac-to-Bac system can be used successfully and rapidly in the expression of viral gene.
Miao
J
J
Department of Hepatitis, Institute of Virology, Chinese Academy of Preventive Medicine, Beijing 100052.
Cong
X
X
Tan
W
W
D016367:Baculoviridae / Q000235:genetics*; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D005822:Genetic Vectors; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis* / Q000235:genetics*; D006515:Hepatitis B virus / Q000235:genetics / Q000276:immunology; D006801:Humans; D008297:Male; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection
D004274:DNA, Recombinant; D006514:Hepatitis B Surface Antigens; D011994:Recombinant Proteins