摘要
采用PCR和酶切的方法获得了HBV的两个基因片段,将它们反向插入真核表达载体pMAMneo和pSV_2dhfr构建成针对HBVs基因和e基因的两个反义RNA真核表达载体pAS_1和pAE_1,以磷酸钙-DNA共沉淀法转染人肝癌细胞系2.2.15,通过ELISA方法检测转染后1～8天内细胞培养液中s抗原和e抗原的表达水平,证实内源性反义RNA可明显地抑制HBV抗原的表达。
Two HBV gene fragments, obtained by PCR and enzyme cutting method, were inserted into eucaryotic expressing vectors pMAMneo and pSV2dhfr in reverse direction to construct two anti-sense RNA eucaryotic expressing vectors-pAS1 and pAE1 which targetting HBV s gene and e gene. Human hepatoma cell line 2. 2- 15 was transfected with pAS1 and pAE1 by calcium phosphate-DNA coprecipitation method. During 1 - 8 days after transfection, the expressing amounts of HB-sAg and HBeAg in the media were tested by ELISA. The studies showed that endogenous antisense RNA could inhibit the expression of HBV antigens markedly.
出处
《天津医药》
CAS
1995年第6期323-325,共3页
Tianjin Medical Journal
关键词
内源性
反义RNA
乙型肝炎病毒
基因表达
endogenous antisense RNA hepatitis B virus (HBV) HBV s gene HBV e gene
