Soluble Expression and Enzyme Activity Analysis of African Swine Fever Virus K196R and A240L Proteins

doi:10.13560/j.cnki.biotech.bull.1985.2020-0216

Abstract

Abstract:

The objective of this work is to provide effective antigens for the clinical diagnosis and immunoprophylaxis of African swine fever virus（ASFV）. K196R and A240L coding sequences derived from ASFV SY18 were executed codon optimization and co-expression with soluble-tag in their N terminals. The target proteins were purified by affinity chromatography after TEV digestion,and their reactivity and enzymatic activity were evaluated through Western blot and enzyme activity detection kit,respectively. As results,the soluble expressions of K196R and A240L proteins were obtained by severally fusing with SUMO and MBP tag,and the size was about 22 kD and 27 kD after tag removed. Western blot results showed both K196R and A240L proteins well specifically reacted with the ASF positive serum. Enzyme activity detection results demonstrated the K196R and A240L proteins all presented definite enzyme catalytic activity in vitro. In conclusion,the soluble K196R and A240L proteins primarily expressed in Escherichia coli all show better reactivity and certain enzymatic activity in vitro,which will be the basis for the ASFV clinical diagnosis and protein relevant functions investigation.

Key words: African Swine Fever, K196R protein, A240L protein, soluble expression, western blot identification, enzymatic activity analysis

LI Peng-hao, LIANG Yan-yu, WANG Yan-wei, GUAN Yang, PANG Wen-qiang, TIAN Ke-gong. Soluble Expression and Enzyme Activity Analysis of African Swine Fever Virus K196R and A240L Proteins[J]. Biotechnology Bulletin, 2020, 36(11): 70-75.

Figures/Tables 5

References 20

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