Apoptosis-related protein Apr.docVIP

 PAGE \* MERGEFORMAT 5 Apoptosis-related protein Apr Author: Li Yuan Fei Shi Jianguo Guo Ailin Guo-Qiang Zhu Qing-Guo Yan [Keywords:] Apr Cloning, sequencing and preliminary expression of apoptosis related protein 2 [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. [Keywords] Apr2 gene; apoptosis; cloning; fusion protein; inclusion bodies [Abstract] Objective: From the HL60 cell apoptosis model of apoptosis-related proteins Apr2 cloned coding region of genes and expressed them, in order to further study the structure and function Apr2 and polyclonal antibody preparation lay the foundation. Methods: HL60 cells, wither death model, HL60 apoptotic cells extracted total RNA, in order to RTPCR method to get Apr2 cDNA coding sequence will be with PGEMT Easy vector, transformation of E.coli DH5α , recombinant cloning vector PGEMTEasy/apr2, sequencing is correct, will the purpose of PGEX4T2 fragment subcloned into the prokaryotic expression vector and transformed into E. coli, IPTG-induced recombinant protein expression, analy