凋亡相关蛋白Apr2的克隆、测序及初步表达
Apr2基因,,Apr2基因;凋亡;克隆;融合蛋白;包涵体,1材料和方法,2结果,3讨论,【参考文献】
Cloning, sequencing and preliminary expression of apoptosis related protein 2LI YuanFei1, SHI JianGuo1, GUO AiLin1, ZHU GuoQiang2, YAN QingGuo1
1Department of Pathology, Xijing Hospital, 2Department of Periodontal Membrane, College of Stomatology, Fourth Military Medical University, Xi’an 710033, China
【Abstract】 AIM: To clone and express the apoptosis related protein 2 (Apr2) from the model of the apoptosis cells of HL60. METHODS: The model of apoptosis cells of HL60 was established, total RNA was isolated from the cells and mRNA was reversely transcribed into cDNA. PCR was used to amplify the apr2 coding region and the PCR product was cloned into PGEMT Easy vector and sequenced. It was then subcloned into expression vector PGEX4T2 and induced with IPTG. RESULTS: Apr2 gene was cloned into PGEMT Easy vector and the sequence was confirmed. SDSPAGE showed that the fusion protein was expressed as inclusion bodies in E. coli. Band density scanning of stained gel was performed to estimate the percentage of the recombinant protein in the total bacteria protein, which was up to 40%. CONCLUSION: Apr2 gene has been successfully cloned and preliminary expression product of fusion protein has been obtained, which lay the basis for further purification and studies of Apr2's structure and function. ......
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