[Construction and biological activity assessment of core mutants of hepatitis B virus adr subtype].
OBJECTIVE
To construct the core mutant (L97 and V60) plasmids of hepatitis B virus (HBV) and assess their biological activity.


METHODS
Site-directed mutagenesis was performed to induce specific core point mutations in HBV adr suhtype 1.2 copy genome plasmid p3.8 II. The plasmids were transfected into HepG2 cells via liposome, and intracellular HBV DNA was analyzed by Southern hybridization, while extracellular HBV antigens (HBsAg and HBeAg) in the culture supernatant were assayed by enzyme-linked immunosorbent assay.


RESULTS
Sequence analysis of the constructed mutant plasmids p3.8 L97 and p3.8 V60 demonstrated mutations at 2 189 A C and 2 086 C HBV adr subtype genome core mutant (L97 and V60) plasmids we have constructed possess biological activity.G respectively. These mutant plasmids exhibited HBV DNA replication activity and gene expression in host cells.


CONCLUSIONS
HBV adr subtype genome core mutant (L97 and V60) plasmids we have constructed possess biological activity.
Chen
Wei-Hong
WH
Department of Infectious Diseases, Nanfang Hospital, First Military Medical University, Guangzhou 510515, China.
Lin
Yu-Long
YL
Wang
Yan-Jun
YJ
Luo
Kang-Xian
KX
D001483:Base Sequence; D003001:Cloning, Molecular; D015967:Gene Expression Regulation, Viral; D006514:Hepatitis B Surface Antigens / Q000378:metabolism; D006513:Hepatitis B e Antigens / Q000378:metabolism; D006515:Hepatitis B virus / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D017354:Point Mutation; D014407:Tumor Cells, Cultured; D014758:Viral Core Proteins / Q000235:genetics*; D014779:Virus Replication / Q000235:genetics
D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens; D014758:Viral Core Proteins