Cloning, sequencing and preliminary expression of apoptosis related protein 2

    LI Yuanª²Fei1, SHI Jianª²Guo1, GUO Aiª²Lin1, ZHU Guoª²Qiang2, YAN Qingª²Guo1

    1Department of Pathology, Xijing Hospital, 2Department of Periodontal Membrane, College of Stomatology, Fourth Military Medical University, Xi¡¯an 710033, China

    ¡¾Abstract¡¿ AIM: To clone and express the apoptosis related protein 2 (Aprª²2) from the model of the apoptosis cells of HLª²60. METHODS: The model of apoptosis cells of HLª²60 was established, total RNA was isolated from the cells and mRNA was reversely transcribed into cDNA. PCR was used to amplify the aprª²2 coding region and the PCR product was cloned into PGEMª²T Easy vector and sequenced. It was then subcloned into expression vector PGEXª²4Tª²2 and induced with IPTG. RESULTS: Aprª²2 gene was cloned into PGEMª²T Easy vector and the sequence was confirmed. SDSª²PAGE 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: Aprª²2 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 Aprª²2's structure and function. ......