Molecular Cloning and Expression of G Protein β Subunit Gene of Curvularia lunata

  • SI He-long ,
  • TONG Ya-meng ,
  • HAO Zhi-min ,
  • LI Zhi-yong ,
  • WANG Nan ,
  • DONG Zhi-ping ,
  • DONG Jin-gao
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  • 1. College of Life Sciences, Agricultural University of Hebei, Baoding 071001, China;
    2. Millet Institute, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang 050035, China;
    3. College of Life Science, Hebei Normal University, Shijiazhuang 050024, China

Received date: 2014-05-12

  Online published: 2014-09-20

Abstract

In order to make a foundation for illustrating the pathogenic mechanism of G protein beta-subunit in Curvularia lunata,this research focuses on acquiring the gene encoding subunit in C.lunata and exploring its expression pattern.The full-length cDNA of G-protein beta-subunit was cloned using SMART RACE RT-PCR,and the expression pattern of ClGβ gene was analyzed under different growth time using Real-time PCR.The full-length cDNA of ClGβ was 1 056 bp and encoded 351 amino acids,while its DNA contained 4 introns and 5 extrons.The cDNA sequence of ClGβ had been deposited in GenBank with accession number JQ768316.The results of Real-time PCR indicated that the gene expression level of G-protein beta-subunit is lower at early stage and higher at later stage.We constructed the prokaryotic expression vector pET28(a)-ClGβ,and E.coli BL21 was transformed by this recombinant construct and induced by IPTG.The molecular weight of the expression product was identical with the calculated molecular weight of ClGβ.

Cite this article

SI He-long , TONG Ya-meng , HAO Zhi-min , LI Zhi-yong , WANG Nan , DONG Zhi-ping , DONG Jin-gao . Molecular Cloning and Expression of G Protein β Subunit Gene of Curvularia lunata[J]. Acta Agriculturae Boreali-Sinica, 2014 , 29(4) : 7 -12 . DOI: 10.7668/hbnxb.2014.04.002

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