新疆野生盘羊ISG15基因的克隆表达

doi:10.11843/j.issn.0366-6964.2013.07.005

Abstract

Abstract:

The aim of this study was to clone and express the ISG15 gene of Xinjiang Wild argali, and to detect the activity of the expression product. The full length cDNA of Xinjiang Wild argali ISG15 gene was cloned by RT-PCR and RACE, then ISG15 gene was cloned into eukaryotic expression vector pPIC9K, the pPIC9K- ISG15 was transformed into Pichia Pastoris GS115 yeast genome by electroporation and the expression was induced by methanol,the expressed protein was purified by Ni²⁺chelate affinity chromatography, finally the activity of the expressed protein was detected by lymphocyte transformation test. The result showed that the full length cDNA of Xinjiang Wild argali ISG15 was 642 bp,the open reading frame was 474 bp, encoded 157 amino acids. The yeast recombinant strains GS115/pPIC9K-ISG15 was used to express ISG15, SDS-PAGE analysis showed the expressed protein was 33 ku, the expressed protein could be purified by Ni²⁺chelate affinity chromatography. The lymphocyte proliferation test results showed that the expressed product could significantly stimulate lymphocyte proliferation(P<0.05), which proved the expressed protein had biological activity.

CLC Number:

S826
S813.3

LU Hai-fu, SHEN Wen, LIU Kai, CUI Ru-peng, YANG Wen, JIANG Fang-pei, SUN Yan-ming. Cloning and Expression of the ISG15 Gene of Xinjiang Wild Argali[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2013, 44(7): 1023-1029.

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Cloning and Expression of the ISG15 Gene of Xinjiang Wild Argali

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