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Expression of hepatitis B virus genes in early embryonic cells originated from hamster ova and human spermatozoa transfected with the complete viral genome

机译:乙型肝炎病毒基因在来源于完整病毒基因组的仓鼠卵和人类精子的早期胚胎细胞中的表达

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摘要

To detect the expression of hepatitis B vims (HBV) genes (HB S and C genes) in early embryonic cells after introducing motile human sperm carrying HBV DNA into zona-free hamster oocytes via the in vitro fertilization (IVF) technique. Methods: Human sperm-mediated HBV genes were delivered into zona-free hamster oocytes by the IVF method. Polymerase chain reaction (PCR) was used to detect HB S and pre-Core/Core (pre-C/C) coding genes both in one- and two-cell embryos. Reverse transcription-PCR (RT-PCR) analysis was used to study the expression of the two genes. Fluorescence in situ hybridization (FISH) analysis using the full-length HBV DNA as the hybridization probe was performed to confirm the integration of viral DNA in the host embryonic genome. Results: Both HB S and pre-C/C coding genes are present and transcribed in one- and two-cell embryos originated from hamster ova IVF with human spermatozoa carrying HBV DNA sequences. Conclusion: Sperm-mediated HBV genes are able to replicate and express themselves in early embryonic cells. These results provide direct evidence that HBV DNA could transmit vertically to the next generation via the male germ line.
机译:在通过体外受精(IVF)技术将携带HBV DNA的运动性人类精子引入无带区的仓鼠卵母细胞后,检测早期胚胎细胞中的乙型肝炎病毒(HBV)基因(HB S和C基因)的表达。方法:通过IVF方法将人类精子介导的HBV基因传递到无透明带的仓鼠卵母细胞中。聚合酶链反应(PCR)用于检测一细胞和两细胞胚胎中的HB S和前Core / Core(pre-C / C)编码基因。逆转录PCR(RT-PCR)分析用于研究这两个基因的表达。使用全长HBV DNA作为杂交探针进行荧光原位杂交(FISH)分析,以确认病毒DNA在宿主胚胎基因组中的整合。结果:HB S和pre-C / C编码基因均存在,并在源自带有人精子的仓鼠卵IVF的一细胞和两细胞胚胎中转录,这些人的精子携带HBV DNA序列。结论:精子介导的HBV基因能够在早期胚胎细胞中复制和表达自身。这些结果提供了直接的证据,证明HBV DNA可以通过雄性种系垂直传播到下一代。

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