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首页> 外文期刊>World Journal of Gastroenterology >Replication of hepatitis B virus in primary duck hepatocytes transfected with linear viral DNA.
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Replication of hepatitis B virus in primary duck hepatocytes transfected with linear viral DNA.

机译：乙型肝炎病毒在线性病毒DNA转染的原代鸭肝细胞中的复制。

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摘要

AIM: To explore the expression and replication of hepatitis B virus (HBV) DNA in primary duck hepatocytes (PDHs). METHODS: Complete HBV genome was transfected into PDHs by electroporation (transfected group, 1.19X10(12) copies of linear HBV DNA/1X10(7) PDHs). After 1-5 d of transfection, HBsAg and HBeAg in the supernatant and lysate of PDHs were measured with the IMX System. Meanwhile, replicative intermediates of HBV DNA were analyzed by Southern blotting and Dot blotting. PDHs electroporated were used as control group. RESULTS: HBsAg in the hepatocyte lysates of transfected group was 15.24 (1 d), 14.55 (3 d) and 5.13 (5 d; P/N values, positive >=2.1) respectively. HBeAg was negative (<2.1). Both HBsAg and HBeAg were negative in the supernatant of transfected group. Dot blotting revealed that HBV DNA was strongly positive in the transfected group and negative in the control group. Southern blot analysis of intracellular total DNA indicated that there were relaxed circular (rc DNA), covalently closed circular (ccc DNA), and single-stranded (ss DNA) HBV DNA replicative intermediates in the transfected group, there was no integrated HBV DNA in the cellular genome. These parameters were negative in control group. CONCLUSION: Expression and replication of HBV genes can occur in hepatocytes from non-mammalian species. HBV replication has no critical species-specificity, and yet hepatic-specific regulating factors in hepatocytes may be essential for viral replication.
机译：目的：探讨乙型肝炎病毒（HBV）DNA在原代鸭肝细胞（PDHs）中的表达和复制。方法：通过电穿孔将完整的HBV基因组转染到PDH中（转染组，线性HBV DNA / 1X10（7）PDHs为1.19X10（12）个拷贝）。转染1-5天后，用IMX系统测量上清液和PDH裂解物中的HBsAg和HBeAg。同时，通过Southern印迹和Dot印迹分析HBV DNA的复制中间体。电穿孔的PDHs用作对照组。结果：转染组肝细胞裂解物中的HBsAg分别为15.24（1 d），14.55（3 d）和5.13（5 d； P / N值，阳性≥2.1）。 HBeAg阴性（<2.1）。转染组上清液中HBsAg和HBeAg均为阴性。点印迹显示，HBV DNA在转染组中呈强阳性，而在对照组中呈阴性。对细胞内总DNA的Southern印迹分析表明，转染组中存在宽松的环状（rc DNA），共价闭合的环状（ccc DNA）和单链（ss DNA）HBV DNA复制中间体，在其中没有整合的HBV DNA。细胞基因组。这些参数在对照组中为阴性。结论：HBV基因的表达和复制可以发生在非哺乳动物的肝细胞中。 HBV复制没有关键的物种特异性，但是肝细胞中的肝特异性调节因子可能对病毒复制至关重要。

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