重组逆转录病毒介导的neo^R基因在公鼠生殖系细胞的转移

Neomycin Resistant Gene Transfer Mediated by Recombinant Retrovirus in Male Mouse Germ Line

将感染滴度为 10 6cfu/ m L、携带新霉素磷酸转移酶 (neo R)基因的重组逆转录病毒与台盼蓝、polybrene按一定比例配制成注射液 ,单侧睾丸注入 14～ 16日龄公鼠曲细精管以体内感染精原干细胞 ,进行重组逆转录病毒 -精原干细胞介导的基因转移研究。注射后 45 d,采集公鼠精液 ,进行精液品质与 PCR检测。结果表明 ,在优化的注射条件下 ,曲细精管内注射重组逆转录病毒后 ,不会干扰小鼠精子的正常发育能力 ,并能够将外源基因整合到精子基因组 ,从而首次通过重组逆转录病毒成功地实现了 neo

In order to study the gene transfer mediated by recombinant retrovirus- spermatogonial stem cells, recombinant retroviruses carrying neo R (titer: 10 6 cfu/ml), together with typan blue and polybrene, were injected into contorted seminiferous tubule of mice(14-16 day) to infect spermatogonial stem cells in vivo. After 45 days, sperms were collected from injected mouse, the sperm quality was examined and PCR technique was employed to examine the integration of neo R into the genome of sperms. The results showed that injection solution did not have influence on the development of sperms,and neo R genes were integrated into the genome of mature sperm. This is the first report that foreign genes of interest have been transferred into male spermatogonial stem cells in vivo by recombinant retrovirus.