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目的了解C基因启动子区T1762A1764变异对其启动子的影响。方法PCR产物直接测序,检测重症乙型肝炎患者乙型肝炎病毒(HBV)前C和C基因调节序列的T1762A1764变异;并将该调节序列插入到氯霉素乙酰转移酶(CAT)表达质粒中,转染HepG2细胞并瞬时表达,比较T1762A1764野株及变异株的C基因启动小区序列及CAT表达水平。结果T1762A1764变异在使CAT表达下调中起主要作用。结论T1762A1764变异使C基因启动子活性下调中起重要作用。
Objective To investigate the effect of mutation of T-1762A1764 in promoter region of C gene on its promoter. Methods The PCR products were directly sequenced to detect the T1762A1764 variation in the regulatory sequences of pre-C and C genes of hepatitis B virus (HBV) in patients with severe hepatitis B. The regulatory sequence was inserted into the expression plasmid of chloramphenicol acetyltransferase (CAT) HepG2 cells were transfected and transiently transfected. The promoter region and CAT expression level of C gene in T1762A1764 wild-type and mutant strains were compared. Results The variation of T1762A1764 played a major role in down-regulation of CAT expression. Conclusion The mutation of T1762A1764 plays an important role in the down-regulation of promoter activity of C gene.