论文部分内容阅读
目的:使去透明带金黄地鼠的卵母细胞与携带乙肝病毒 DNA(HBV DNA)的人类精子体外受精,检测乙肝病毒基因(S 基因与 C 基因)在早期胚胎细胞中的复制与表达。方法:通过异种体外受精,由人类精子将乙肝病毒基因导入去透明带地鼠卵母细胞中;用聚合酶链反应(PCR)检测单细胞和2-细胞胚胎细胞的基因组中是否有 S 基因和前 C/C 基因;用逆转录聚合酶链反应(RT-PCR)研究上述基因是否在胚胎细胞中表达。以全长 HBV DNA 制备探针,与胚胎细胞制片进行荧光原位杂交(FISH),观察胚胎细胞基因组中是否有 HBV DNA整合。结果:PCR、RT-PCR 和 FISH 分析均在待测样本中获得阳性结果。结论:以精子为载体、携带到卵内的 HBV 基因能够在早期胚胎细胞中复制和表达,该结果为 HBV 有可能通过男性生殖细胞垂直传递给子代提供了直接证据。
OBJECTIVE: To in vitro fertilize the oocytes of golden hamster with human sperm carrying HBV DNA and to detect the replication and expression of hepatitis B virus (S and C) genes in early embryonic cells. Methods: HepG2 gene was introduced into zona pellucida oocytes by human sperm by in vitro in vitro fertilization. The S gene was detected by polymerase chain reaction (PCR) in the genomes of single-cell and 2-cell embryos Pre-C / C gene; Reverse transcription-polymerase chain reaction (RT-PCR) was used to investigate whether these genes were expressed in embryonic cells. Probes were prepared from full-length HBV DNA, and fluorescence in situ hybridization (FISH) was performed with embryonal cells to see if HBV DNA integration was present in the embryonic cell genome. Results: Positive results were obtained in the samples tested by PCR, RT-PCR and FISH analysis. CONCLUSIONS: The sperm-borne HBV gene can be replicated and expressed in early embryo cells. This result provides direct evidence that HBV may be delivered vertically to male offspring through male germ cells.