包头医学院学报
包頭醫學院學報
포두의학원학보
JOURNAL OF BAOTOU MEDICAL COLLEGE
2015年
5期
1-3
,共3页
胶质细胞源性神经营养因子%表达载体%STO细胞%转基因
膠質細胞源性神經營養因子%錶達載體%STO細胞%轉基因
효질세포원성신경영양인자%표체재체%STO세포%전기인
GDNF%Expression vectors%STO cells%Gene transfer
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。
目的：製備過錶達GDNF的轉基因 STO 細胞，用于富集精原榦細胞。方法：構建 GDNF、EGFP 雙順反子錶達載體，採用XhoⅠ、SspⅠ酶切法鑒定重組質粒，用脂質體介導法將質粒轉入STO細胞，經G418抗性篩選後，分離擴增培養錶達綠色熒光蛋白的抗性細胞，採用PCR法鑒定轉基因細胞。結果：成功構建重組GDNF錶達載體，將其轉染STO細胞後，能夠穩定整閤到細胞染色體中，穫得瞭錶達綠色熒光蛋白的轉GDNF基因STO細胞。結論：通過構建錶達載體，製備GDNF轉基因STO細胞，為富集精原榦細胞的研究奠定瞭基礎。
목적：제비과표체GDNF적전기인 STO 세포，용우부집정원간세포。방법：구건 GDNF、EGFP 쌍순반자표체재체，채용XhoⅠ、SspⅠ매절법감정중조질립，용지질체개도법장질립전입STO세포，경G418항성사선후，분리확증배양표체록색형광단백적항성세포，채용PCR법감정전기인세포。결과：성공구건중조GDNF표체재체，장기전염STO세포후，능구은정정합도세포염색체중，획득료표체록색형광단백적전GDNF기인STO세포。결론：통과구건표체재체，제비GDNF전기인STO세포，위부집정원간세포적연구전정료기출。
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, SspⅠenzyme 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 ex-pression 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 .
