摘要
目的定量观察双位点核酶对细胞内ＨＢＶ基因表达的抑制作用。方法构建针对ＨＢＶＣ区双位点核酶的真核表达载体（ｐＢＢＳ２１２－Ｒｚ），将该质粒与ｐｌ．２Ⅱ（含ＨＢＶ全序列），共转染人肝细胞癌（ＨＨＣＣ）细胞株，对转染细胞所表达ＨＢｅＡｇ进行间接免疫荧光染色，用共聚焦显微镜对其含量进行定量分析。结果共转染细胞可以表达ＨＢｅＡｇ，同对照组相比，核酶组荧光量明显下降。结论在核酶与ＨＢＶ共转染细胞中，核酶通过对ＨＢＶｍＲＮＡ的剪切作用，使ＨＢｅＡｇ的表达量明显受到抑制。
Objective To study the inhibition of HBeAg
expression in cells by twounit ribozyme. Methods Plasmid pBBS212-R2,a eukaryotic
expression vector with a twounit ribozyme against hepatitis B virus, was constructed and
cotransfected with p1.2 plasmid (carring genome of adrsubtype HBV) in to the HHCC cell
(Human Hepatocelluar Carcinoma cell line). Immunoflurescence tests were applied to detecte
the cotransfected cells, and data was analysed with confocal microscope quantitively. Results
The cotransfected cells expressed HBeAg, while the quantity of fluorecein was reduced
significantly in comparison with control cells. Conclusion Ribozyme can inhibit the expression
of HBeAg in the cotransfected cells by cleaving the mRNA of HBV.
出处
《中华传染病杂志》
CAS
CSCD
北大核心
1999年第3期152-154,共3页
Chinese Journal of Infectious Diseases
基金
国家自然科学基金
