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人akt1基因真核表达载体的构建及表达
http://www.100md.com 《第四军医大学学报》 2005年第8期
akt1基因,,akt1基因;,豆蔻酰化;,逆转录聚合酶链反应;,克隆;,基因表达,1材料和方法,2结果,3讨论,【参考文献】
     Construction and expression of human akt1 gene

    SHEN Lan, LI Xia, ZHANG ZiFeng, SU Jin, LIU XinPing, YAO LiBo

    Department of Biochemstry and Molecular Biology, School of Basic Medicine, Fourth Military Medical University, Xian 710033, China

    【Abstract】 AIM: To construct the eukaryotic expressing vectors of human akt1 and Nterminally myristoylation signalattached akt1 (myrakt1) and to test their expression in African green monkey kidney cell line COS7. METHODS: Human akt1 and myrakt1 genes tagged with flag were amplified from human dental pulp mRNA by RTPCR. The products were cloned into pMD18T vector, sequenced and then subcloned into eukaryotic expressing vector pcDNA3.1(+). The recombined vectors were then transfected into COS7 cells with lipofectin to detect the expression of akt1 and myrakt1 by Western blot. The cell cycle profiles of COS7 cells were analyzed by flow cytometry (FCM). RESULTS: The eukaryotic expressing vectors containing human akt1 and myrakt1 were successfully constructed and their expression in COS7 cells could be detected. AKT1 overexpression and activation showed little effect on the cell cycle of COS7 by cell cycle profiles. CONCLUSION: The eukaryotic expressing vectors containing human akt1 and myrakt1 are constructed and they can express flagakt1 and flagmyrakt1 fused proteins correctly in COS7 cells. But the regulation of cell cycle by AKT1 has to be further investigated. ......

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