牛α0干扰素在毕赤酵母中的分泌表达及抗病毒活性鉴定

doi:10.11843/j.issn.0366-6964.2015.01.016

Abstract

Abstract:

In this study，we cloned BoIFN-α0 mature peptide gene and expressed it in the eukaryocyte， and also studied the activity of this protein.To achieve secretary expression of bovine interferon-α0 (BoIFN-α0 ) in Pichia pastoris，the BoIFN-α0 mature peptide gene was synthesized by PCR and inserted into the secreted expression vector pPICZαA.The Pme Ⅰ linearized resultant recombinant plasmid was transformed into P.pastoris GS115 by electroporation and the positive recombinant strains P.pastoris were selected by 100 μg•mL^-1 of zeocin^TM and identified by PCR with the AOX1 primers and the specific primers.The positive recombinant strains were induced with methanol and achieved secretory expression in P.pastoris Yeast expression system.The products in culture medium were detected by SDS-PAGE and Western blot.The SDS-PAGE results showed that the recombinant protein was expressed in the supernatant with molecular of 18 and 21 kDa (with the difference of glycosylation).Furthermore，the BoIFN-α0 was able to efficiently inhibiting virus replication in vesicular stomatitis virus (VSV)/MDBK cells detection system and its antiviral activity was about 5.72×10⁶ U•mg^-1.These results provided a basis for further application of BoIFN-α.

CLC Number:

S852.4

PENG Tong-quan，SHAO Jian-wei，ZHANG Run-xiang，CAO Chong，MA Bo，GAO Ming-chun，WANG Jun-wei. Expression of the Bovine Interferon Alpha0 with Antiviral Activity in Pichia Pastoris[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(1): 124-129.

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Expression of the Bovine Interferon Alpha0 with Antiviral Activity in Pichia Pastoris

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