中文摘要：

目的从昆明鼠睾丸中克隆Bmi1基因,构建真核表达载体,并转染支持细胞,以便用作培养精原干细胞（SSCs）的滋养层.方法以5日龄昆明鼠为材料,提取小鼠睾丸组织中总RNA后,以RT-PCR技术克隆小鼠睾丸Bmi1基因,构建真核表达载体,并转染TM4细胞（睾丸支持细胞株）,在转染后40 h进行免疫荧光鉴定.结果成功克隆小鼠睾丸Bmi1基因的cDNA,测序正确;免疫荧光细胞染色显示,转染后的支持细胞中有Bmi1蛋白表达.结论本研究为以转染了Bmi1基因的支持细胞作饲养层培养SSCs奠定了基础.

英文摘要：

Objecave：To clone Bmi1 gene from Kunming mouse testis,construct the eukaryotic expression vector and transfect this vector into Sertoli cells in order to use the Bmi1-transfected Sertoli cells as the feeder layer to cultivate sperrnatogonial stem cells（SSCs）.Methods：Total RNA was extracted from the testes of 5 day old Kunming mice and Bmi1 was cloned and amplified using RT-PCR,inserted into the eukaryotic expression vector and transfected into sertoli cells（TM4 cell line）.Immunofluorescence with anti-Bmi1 antibodies was performed at 40 h following the transfection.Results：Bmi1 DNA was cloned successfully,and Bmi1 expressed at 40 h after transfected into Sertoli cells.Conclusion：The present study provides a basis for culturing SSCs with Bmi1-transfected sertoli cells as the feeder layer.