论文部分内容阅读
目的 体外研究乙型肝炎病毒Ｃ（ＨＢＶ／Ｃ） 基因变异的生物学意义。方法 应用逆转录病毒载体ｐＸＴ１ 构建ＨＢＶ／Ｃ 基因表达载体，并转染永生化人外周血Ｂ 细胞使之稳定表达ＨＢｃＡｇ。结果 重组质粒ｐＸＴ１ＨＢＶ／Ｃ 经聚合酶链反应（ＰＣＲ） 和ＢｇｌⅡ及ＸｈｏⅠ双酶切鉴定均阳性，转染后的永生化人外周血Ｂ 细胞ＰＣＲ 鉴定含目的ＤＮＡ，流式细胞仪分析显示，约４７ ．４ ％ 的细胞膜上表达了ＨＢｃＡｇ。结论 ＨＢＶ／Ｃ基因逆转录病毒表达载体有较高的转染效率，目的基因能在宿主细胞中稳定表达，有利于系列研究ＨＢＶ／Ｃ基因变异的生物学意义。
Objective To study the biological significance of hepatitis B virus (HBV / C) gene mutation in vitro. Methods The HBV / C gene expression vector was constructed by retroviral vector pXT1 and transfected into immortalized human peripheral blood B cells to stably express HBcAg. Results The recombinant plasmid pXT1HBV / C was identified by polymerase chain reaction (PCR) and restriction endonuclease digestion with BglⅡ and XhoⅠ. The transfected immortalized human peripheral blood B cells were identified by PCR and identified by flow cytometry , About 47. 4% of the cell membrane expressed HBcAg. Conclusion The retroviral vector of HBV / C gene has high transfection efficiency and the target gene can be stably expressed in host cells, which is beneficial to study the biological significance of HBV / C gene mutation in series.