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HEK293细胞中CDK10蛋白复合体的分离纯化 被引量：1

Isolation and Purification of the CDK10 protein complex from HEK293 cell
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摘要 通过半定量RT-PCR(reverse transcriptase PCR)检测了CDK10在人胚胎肾细胞(HEK293)、人宫颈癌细胞(HeLa)、人乳腺癌细胞(MCF7)中的表达水平,克隆并构建了全长CDK10基因的真核表达质粒.在HEK293中筛选得到了稳定表达CDK10-Flag的细胞系,大规模培养该细胞系,分离细胞核、细胞质提取物,在非变性条件下利用anti-Flag抗体免疫共沉淀(Immunoprecipitation,IP),银染及蛋白质免疫印迹检测结果表明分离到CDK10蛋白及与其可能结合的其它蛋白,为质谱(Mass Spectrum,MS)鉴定提供材料. The expression of Cdk10 was checked in HEK293 HeLa and MCF7 cells using Semi Quantita- tive RT-PCR. Plasmid pcDNA3, 1（-）-Cdk10-Flag was constructed after cloning of Cdk10. HEK293 cell lines that can stably express Cdk10-Flag were cultured in large numbers. Cdk10 complex was puri- fied from cell extractions using anti-Flag immunoprecipitation under native conditions. The purified Cdk10 complex paved the way for future MS identification.
作者 赵长宏 陈斌 吴传芳 欧阳劲 秦岭
机构地区 四川大学生命科学学院 成都南诺格生物科技有限责任公司 成都华高瑞甜科技有限公司
出处 《四川大学学报（自然科学版）》 CAS CSCD 北大核心 2013年第1期187-192,共6页 Journal of Sichuan University(Natural Science Edition)
基金 国家自然科学基金(30970625)
关键词 CDK10 蛋白纯化 细胞周期蛋白依赖性蛋白激酶 银染 CDK10, protein purification, cyclin-dependent kinases,silver stain
分类号 Q26 [生物学—细胞生物学]
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参考文献12

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同被引文献14

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四川大学学报（自然科学版）
2013年 第1期

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