Prokaryotic Expression of PPRV Nigeria75/1 H Protein and Prediction of Antigen Epitope

doi:10.7668/hbnxb.2018.05.016

Abstract

Abstract: To clone the gene encoding hemagglutinin(H)protein of Nigeria 75/1 strain of attenuated Peste des petits ruminants virus(PPRV),express in prokaryotic cells and prepare its polyclonal antibody by bioinformatics. According to the encoding sequence of PPRV H gene(GenBank X74443),a pair of primers were designed,with which the overall length were amplified by RT-PCR and inserted into vector pET-32a. The constructed recombinant plasmid was transformed to E. coli Rosetta for expression under the induction of 1 mmol/L IPTG at 37℃. The expressed products were purified by affinity chromatography using the nickel ion protein purification volume with His tag,and identified by Western Blot. Preparing its polyclonal antibody by bioinformatics.Restriction analysis proved that recombinant plasmid pET-32a-H was constructed correctly. The expressed PPRV H protein,with a relative molecular mass of about 67 ku. The prepared polyclonal antibody recognized the whole viral antigen of PPRV and recombinant PPRV H protein expressed in baculovirus. Recombinant plasmid pET-32a-H was constructed successfully,and PPRV H protein was successfully expressed. Useing bioinformatics could find polyclonal antibody 5-8,14-16,73-75,83-90,125-131,142-147,170-177,236-245,281-285,312-317,360-363,370-379, 388-391,445-449,487-489,503-505,520-522,532-535,544-551,592-595. The positive plasmid pET-32a-H was successfully constructed,and the target protein was expressed. The potential epitope of H protein was successfully predicted. It is important for elimination of PPRV earlyer and preparation of novel subunit vaccines.

Key words: Peste des petits ruminants virus, H protein, Prokaryotic expression, B cell epiyope

CLC Number:

LI Linjie, CHANG Qiuyan, MA Peng, WANG Yueying, MA Xiaoxia, BAI Jialin. Prokaryotic Expression of PPRV Nigeria75/1 H Protein and Prediction of Antigen Epitope[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2018, 33(5): 111-116. doi: 10.7668/hbnxb.2018.05.016.

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