论文部分内容阅读
目的 构建HBVX蛋白真核表达质粒并了解其对反式转录激活作用的影响。方法 用亚克隆方法构建HBVX蛋白真核表达质粒 ,以氯霉素乙酰转移酶转化率分析 (CAT试验 )揭示HBX蛋白的反式转录激活作用。结果 HBVX蛋白真核表达质粒转染HepaG2细胞后 ,以WesternBlot检测发现 2 μg、5 μg和 10 μg的 p5SGUTPLHBXDNA均能有效表达 ;CAT试验揭示 2 μg、5 μg、10μg的 p5SGUTPLHBXDNA对报道基因 pHE2×CAT的14 C -标记乙酰化氯霉素转化率分别为 (4 5 .5± 2 5 .9) %、(6 5 .6± 14.4) %和 (6 8.8± 19.7) %。结论 本结果为进一步研究HBVX蛋白的反式转录激活作用打下了基础
Objective To construct the eukaryotic expression vector of HBVX protein and study its effect on transactivation. Methods The eukaryotic expression plasmid of HBVX protein was constructed by subcloning method. The transactivation of HBX protein was revealed by the CAT assay of chloramphenicol acetyl transferase activity. Results After transfection of HepG2 cells with eukaryotic expression vector of HBVX protein, the results showed that 2 μg, 5 μg and 10 μg of p5SGUTPLHBX DNA were efficiently expressed by Western Blot. The results of CAT assay revealed that the expression of pEVSGUTPLHBX2, 2 μg, 5 μg and 10 μg, The 14 C - labeled acetylated chloramphenicol conversion rates were (45.5 ± 25.5)%, (65.5 ± 14.4)% and (6.48 ± 19.7)%, respectively. Conclusion The results laid the foundation for further study of transactivation of HBVX protein