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Clone of Tams1 Gene Non-hydrophobic Region of Theileria annulata and High Level Expression of Recombinant Protein GST-P27

曹雯丽1,王真1,沙它尔·卡哈尔1,2,王冰洁1,巴音查汗1   

  1. (1.College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, China;2.Animal Husbandry and Veterinary Station of Turpan City, Turpan 838000, China)
  • Received:2013-08-07 Online:2014-03-20 Published:2014-05-15

Abstract: A fragment about 696 bp was amplified by PCR technique with specific primers based on Tams1 gene sequence(AF214842) of Theileria annulata reported in GenBank. Then the target fragment was directionally cloned into pGEX-4T-2 expression vector, the recombinant plasmid DNA was cut by enzymes, and then sequenced.The result showed that the homology of the cloned Tams1 gene was 98%. The positive plasmid was transformed into BL21(DE3), and then induced by IPTG. Soluble fusion protein whose expression level reached 1.39 mg/mL was obtained when it was induced with 0.5 mmol/L IPTG for 4 h at 37 ℃, and it was 53 ku. Western blotting showed that recombinant protein GST-P27 which was purified by Glutathione Sepharose 4B had the favorable reactionogenicity.

Key words: Theileria annulata; Tams1 gene; clone; GST-P27; high level expression