Cloning, eukaryotic expression of human ISG20 and preliminary study on the effect of its anti-HBV.
Human ISG20 gene was cloned and the effect of its anti-HBV was primarily studied. The ISG20 gene was amplified from HeLa cells by RT-PCR and recombinant vector expressing ISG20 was constructed by genetic engineering. The overexpression of ISG20 in HepG2 cells was detected by Western blot and the levels of secretion of HBs antigen and HBe antigen tested by ELISA. The results showed that: (1) Sequence of ISG20 cloned was consistent to that published in Genebank; (2) Recombinant vector expressing ISG20 could be expressed in HepG2 cells by transfection; (3) The overexpression of ISG20 protein could reduce the levels of the secretion of HBs antigen and HBe antigen in transfected HepG2 cells. It was suggested that the overexpression of recombinant ISG20 in culture cells could reduce the synthesis of HBV proteins.
Hao
Youhua
Y
Division of Clinical Immunology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China. youhuahao@hotmail.com
Yang
Dongliang
D
D045744:Cell Line, Tumor; D049109:Cell Proliferation; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D005092:Exonucleases / Q000096:biosynthesis / Q000235:genetics*; D005095:Exoribonucleases; D005786:Gene Expression Regulation; D015967:Gene Expression Regulation, Viral; D005818:Genetic Engineering / Q000379:methods; D006367:HeLa Cells; D006509:Hepatitis B / Q000150:complications / Q000628:therapy; D006515:Hepatitis B virus / Q000235:genetics*; D022781:Hepatocytes / Q000378:metabolism / Q000821:virology; D006801:Humans; D011994:Recombinant Proteins / Q000737:chemistry
D011994:Recombinant Proteins; D005092:Exonucleases; D005095:Exoribonucleases; C112346:ISG20 protein, human