The Fusion-expression of Wheat TaNADP-ME1gene in E．coli and Purification of Fusion Protein

doi:10.3969/j.issn.1000-7091.2013.02.001

Abstract

Abstract: NADP-dependent malic enzyme(NADP-ME)is a key enzyme in C4photosynthesis,the objective is to construct the TaNADP-ME1gene into prokaryotic expression vector,express fusion protein in E. coli and purify the fusion protein. The TaNADP-ME1gene was constructed into expression vector pET32a by recombination technolo-gy,recombination plasmid was identified by digestion with restriction enzymes and PCR amplification,and trans-formed into BL21(DE3)pLysS by CaCl2method,fusion protein was induced by IPTG and purified by Ni2+-NTA agarose column. This research successfully acquired the recombination vector pETE1,and TaNADP-ME1gene was accurately expressed in BL21(DE3)pLysS,SDS-PAGE revealed that the molecular weight of purified fusion protein was about 80 kDa and successfully acquired the fusion protein. This study laid agood foundation for identification thegene function of TaNADP-ME1gene.

Key words: Wheat, TaNADP-ME1gene, Fusion expression, Protein purification

CLC Number:

FU Zhen-yan, ZHANG Zheng-bin, WANG Xiao-jun, XU Ping. The Fusion-expression of Wheat TaNADP-ME1gene in E．coli and Purification of Fusion Protein[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2013, 28(2): 1-5. doi: 10.3969/j.issn.1000-7091.2013.02.001.

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