人akt1基因真核表达载体的构建及表达
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 geneSHEN Lan, LI Xia, ZHANG ZiFeng, SU Jin, LIU XinPing, YAO LiBo
Department of Biochemstry and Molecular Biology, School of Basic Medicine, Fourth Military Medical University, Xian 710033, China
【Abstract】 AIM: To construct the eukaryotic expressing vectors of human akt1 and Nterminally myristoylation signalattached akt1 (myrakt1) and to test their expression in African green monkey kidney cell line COS7. METHODS: Human akt1 and myrakt1 genes tagged with flag were amplified from human dental pulp mRNA by RTPCR. The products were cloned into pMD18T vector, sequenced and then subcloned into eukaryotic expressing vector pcDNA3.1(+). The recombined vectors were then transfected into COS7 cells with lipofectin to detect the expression of akt1 and myrakt1 by Western blot. The cell cycle profiles of COS7 cells were analyzed by flow cytometry (FCM). RESULTS: The eukaryotic expressing vectors containing human akt1 and myrakt1 were successfully constructed and their expression in COS7 cells could be detected. AKT1 overexpression and activation showed little effect on the cell cycle of COS7 by cell cycle profiles. CONCLUSION: The eukaryotic expressing vectors containing human akt1 and myrakt1 are constructed and they can express flagakt1 and flagmyrakt1 fused proteins correctly in COS7 cells. But the regulation of cell cycle by AKT1 has to be further investigated. ......
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