犬α-干扰素蛋白在不同表达载体中的生物学活性比较

doi:10.16431/j.cnki.1671-7236.2016.10.008

Abstract

Abstract:

According to the sequence of canine interferon-α (CaIFN-α) gene published in GenBank,primers were designed,synthesized and introduced EcoRⅠand HindⅢ restriction sites.Using canine liver DNA as a template,CaIFN-α gene was amplified by PCR,then the gene was cloned into pBV220 and pET-32a(+) expression vector,and transformed into the BL21(DE3) expression system.Recombinant fusion protein was analysed by SDS-PAGE and Western blotting.Protein was purified and the antiviral activity was detected by MDCK/VSV cytopathic inhibition assay.The results showed that pBV220-α-IFN recombinant protein has a low activity,but the activity of pET32a-α-IFN recombinant protein was up to 2.56×10⁶ U/mL.This study laid a foundation for production of highly active IFN.

Key words: canine; interferon-α; prokaryotic expression; antiviral activity

CLC Number:

LIU Huan, JIN Hong-yan, HOU Qiang, SHI Qiu-mei, DONG Rui-kai, ZHANG Tong-ming, HOU Shao-hua, SHEN Ping. The Comparison of Canine α-Interferon Antiviral Activity in Different Expression Vectors[J]. , 2016, 43(10): 2560-2565.

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The Comparison of Canine α-Interferon Antiviral Activity in Different Expression Vectors

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