Cloning, prokaryotic expression and protein separation of 6R, an Andrias davidianus ranaviurs (ADRV) infection-related gene

Cloning, prokaryotic expression and protein separation of 6R, an Andrias davidianus ranaviurs (ADRV) infection-related gene
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Affiliation:1. State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China; 2. University of Chinese Academy of Sciences, Beijing 100049, China
Clc Number:S94
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gene was. Then, the recombinant prokaryotic expression plasmid pET-32a-6R was constructed and sequenced. ADRV gene was blasted with homologous sequences from other rana viruses of aquatic animals. The gene encodes a protein of 236aa with a predicted molecular mass of 27 kD. Amino acid sequence alignment of ADRV members in the viruses-6R was transformed into DE3 and induced by IPTG. The expressed protein was purified with Ni-NTA His-Bind affinity chromatography, separated by different concentrations of imidazole eluent and analyzed by SDS-PAGE. A 45 kD fusion proteincontaining protein and 18 kD his-tagged was obtained in accordance with the predicted molecular weight. These results provide useful experimental material for antibody preparation and study on ADRV-host interactions.

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李涛,陈中元,张奇亚. 一个与大鲵蛙病毒 (ADRV) 感染相关基因—6R的克隆、原核表达及其产物分离[J]. Jounal of Fishery Sciences of China, 2015,[volume_no](2):353-358

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Online: June 29,2015
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