CONSTRUCTION OF EUKARYOTIC EXPRESSION VECTORS WITH HOT SPOT MUTATIONS IN HBV SIGNAL PEPTIDE REGION
- VernacularTitle:HBV信号肽区热点变异真核表达载体的构建及对HBeAg表达的影响
- Author:
Yulong LIN
;
Jinlin HOU
;
Zhanhu WANG
- Publication Type:Journal Article
- Keywords:
site mutagenesis;
eukaryotic expression vector;
transfection
- From:
Medical Journal of Chinese People's Liberation Army
2001;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
Molecular biological methods were used to construct the eukaryotic expression vectors of precore and core gene named EBO PreC/C. Then T1862, A1896, A1899 and A1896+A1899 variants were constructed by site mutagenesis in vitro. By PCR RFLP and sequencing ,T1862,A1896,A1899 and A1896+A1899 variants were obtained. Wild type and variants were transfected to HepG2 cells, and HBeAg was tested to observe the difference of HBeAg expression between wild type and variants. After stable expression in HepG2 cells, HBeAg was detected to be positive in cells transfected wild type and A1899 variants, and negative in cells transfected with T1862, A1896,A1896+A1899 variants. The construction of these variants will play an important pole in studying the relation of PreC/C mutations and HBV expression and replication of HBV genome.
