GDNF表达载体的构建及其转染STO细胞的研究  

Construction of the GDNF expression vector and study on the transfected STO cells

作　　者：郭永芝[1] 张学明[2] 

机构地区：[1]包头医学院第一附属医院,内蒙古包头014010 [2]包头医学院基础医学与法医学院

出　　处：《包头医学院学报》2015年第5期1-3,共3页Journal of Baotou Medical College

基　　金：国家自然科学基金(31060304);内蒙古高等学校科学研究项目(NJ10184)

摘　　要：目的:制备过表达GDNF的转基因STO细胞,用于富集精原干细胞。方法:构建GDNF、EGFP双顺反子表达载体,采用XhoⅠ、SspⅠ酶切法鉴定重组质粒,用脂质体介导法将质粒转入STO细胞,经G418抗性筛选后,分离扩增培养表达绿色荧光蛋白的抗性细胞,采用PCR法鉴定转基因细胞。结果:成功构建重组GDNF表达载体,将其转染STO细胞后,能够稳定整合到细胞染色体中,获得了表达绿色荧光蛋白的转GDNF基因STO细胞。结论:通过构建表达载体,制备GDNF转基因STO细胞,为富集精原干细胞的研究奠定了基础。Objective： To prepare transgenic STO cells of over- expressing GDNF which could be used to enrich spermatogonial stem cells.Methods： GDNF and EGFP bicistronic expression vectors were constructed; the recombinant plasmid was identified by using XhoⅠ,SspⅠ enzyme digestion methods and was transfected into STO cells by using liposome mediated methods. After selection with G418,resistant cells expressing green fluorescence protein were isolated,cultured and expanded; the transgenic cells were identified by PCR. Results： The recombinant GDNF expression vector was successfully constructed and integrated into the cell chromosomes after being transfected into STO cells,GDNF transgenic STO cells expressing green fluorescence protein obtained. Conclusion： Construction of expression vectors and preparation of GDNF transgenic STO cells lay a foundation for the research of enriching spermatogonial stem cells.

关 键 词：胶质细胞源性神经营养因子 表达载体 STO细胞 转基因 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]