贾三三;李婷;白欣艳;温丽敏;王玉晶;王海龙;解军;郭睿;

• 1:山西医科大学生物化学与分子生物学教研室
• 2:山西医科大学寄生虫学教研室

摘要(Abstract)：

目的构建小鼠丝氨酸/精氨酸蛋白特异激酶2(serine/arginine-rich protein specific kinase 2,SRPK2)基因真核表达质粒,建立稳定过表达SRPK2的He La细胞系,并探讨SRPK2过表达对He La细胞增殖和凋亡的影响。方法利用RT-PCR从小鼠睾丸组织中扩增SRPK2基因,与p3×Flag-CMV-14质粒连接,构建重组真核表达质粒p3×Flag-CMV-14-SRPK2,转染He La细胞,通过G418筛选获得稳定转染细胞系。采用RT-PCR检测稳定转染He La细胞系中SRPK2基因的转录,Western blot检测SRPK2的表达;MTT法和流式细胞术分别检测SRPK2过表达对He La细胞增殖和凋亡的影响。结果菌落PCR、双酶切及DNA测序表明重组真核表达质粒p3×Flag-CMV-14-SRPK2构建正确;RT-PCR和Western blot分析表明SRPK2基因在He La细胞中稳定过表达;p3×Flag-CMV-14-SRPK2稳定转染组细胞增殖活力明显高于p3×Flag-CMV-14转染组和未转染组(P<0.05),细胞凋亡率明显低于上述两组(P<0.05)。结论成功构建了SRPK2基因的真核表达质粒,建立了稳定过表达SRPK2的He La细胞系,SRPK2过表达可增强He La细胞的增殖活力,而抑制其凋亡。

关键词(KeyWords)： 丝氨酸/精氨酸蛋白特异激酶2;HeLa细胞;细胞增殖;细胞凋亡

Abstract:

Keywords:

基金项目(Foundation): 山西省回国留学人员科研资助项目(2011-043)

作者(Author): 贾三三;李婷;白欣艳;温丽敏;王玉晶;王海龙;解军;郭睿;

Email:

DOI: 10.13200/j.cnki.cjb.001317

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