Cloning,Expression and Bioinformatics Analysis of Maize Zmcen Gene

doi:10.7668/hbnxb.2016.03.003

Abstract

Abstract: To study the characteristics and biological function of Zmcen gene,investigate the protein structure of Zea mays L.,the ORF of maize Zmcen gene was cloned into the expression vector pGEX-6p-1 through PCR method,and the GST recombined expression vector pGEX-6p-Zmcen was constructed.Recombinant GST-ZmCen fusion protein was expressed as soluble protein in Escherichia coli BL21 (DE3) after induction with 0.3 mmol/L IPTG at 27 ℃ for 20 h.The ZmCen protein with the GST affinity chromatography and verified the protein by 12% SDS-PAGE and Western Blotting with anti-GST antibody were purified.Fusion protein was cut out the GST-Tag by PPase and was purified.And by the method of biological information,the results showed that the Zmcen gene was located on the 7^# chromosome,and the full-length gene had 6 exons and 7 introns.The ZmCen protein domain search and alignment analysis were conducted by the maizeGDB resources,and the ZmCen was EF-hand domain family protein.The CDS region of Zmcen gene was 519 bp,encoding 172 amino acid with a molecular weight of 19.78 kDa and pI of 4.78.Using maizeGDB database searching,the expression level analysis showed that Zmcen gene expression had a higher level in cells divided vigorous of the entire maize life cycle.The evolutionary history of centrins was analyzed by means of phylogenetic analysis in plants.The centrin gene was a highly 80.21% homology among centrins of 16 different plants.The results of the research can provide a theoretical basis for exploring the unknown biological function of ZmCen protein.

Key words: Zmcen gene cloning, Expression, Purification, Bioinformatics

CLC Number:

LEI Haiying, BAI Fenglin, LIU Jianxia, WANG Zhijun. Cloning,Expression and Bioinformatics Analysis of Maize Zmcen Gene[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2016, 31(3): 18-24. doi: 10.7668/hbnxb.2016.03.003.

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