中文摘要：

通过半定量RT-PCR（reversetranscriptasePCR）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（MaSSSpectrum，MS）鉴定提供材料．

英文摘要：

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.