小鼠Bmi1基因的克隆及其在支持细胞中的表达  

Cloning of Bmi1 DNA from Mouse Testis and Its Expression in Sertoli Cell

作　　者：郭玉萍[1] 赵洪涛[1] 王玮[1] 张世庆[2] 李澜[2] 李恩中[3] 李德雪[2] 

机构地区：[1]安阳师范学院体育系,河南安阳455002 [2]军事医学科学院基础医学研究所,北京100850 [3]黄淮学院生物工程系,河南驻马店463000

出　　处：《天中学刊》2011年第5期7-9,共3页Journal of Tianzhong

基　　金：国家自然科学基金资助项目(30972090);河南省自然科学基金资助项目(102300410036);河南省高校科技创新人才资助计划(2011HASTIT031);河南省高校青年骨干教师资助计划;黄淮学院青年骨干教师资助计划;总后卫生项目(08ZL030)

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.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.

关 键 词：Bmi1基因 转染 小鼠支持细胞 

分 类 号：Q492.4[生物学—生理学] Q78