Inducible Expression on Multi-epitope of Porcine Circovirus Type 2 and Its Immunological Competence

In order to obtain induction expressed porcine circovirus type 2（PCV2）multi-epitope with good immunogenicity in vitro.Major epitopes of PCV2 were screened in the test,epitopes were composed sequentially,the c DNA sequences were artificially synthesized.Bam HⅠand SalⅠwere directionally cloned into pr...

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Bibliographic Details
Published in动物与饲料科学：英文版 Vol. 8; no. 3; pp. 151 - 154
Main Author Dong Lin Wang Yanping Wang Jinliang Mo Ling Shen Zhiqiang Liu Zengshan
Format Journal Article
LanguageEnglish
Published 2016
Subjects
PCV2
SDS-PAGE
sds-page分析
免疫能力
多表位
猪圆环病毒2型
蛋白多肽
诱导表达
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Summary:In order to obtain induction expressed porcine circovirus type 2（PCV2）multi-epitope with good immunogenicity in vitro.Major epitopes of PCV2 were screened in the test,epitopes were composed sequentially,the c DNA sequences were artificially synthesized.Bam HⅠand SalⅠwere directionally cloned into prokaryotic expression vector PEGX-4T-1 multiple cloning site,then BL21 competent cells were transformed,positive clones were screened,IPTG inducible expression was conducted.Expression on target gene was analyzed by SDS-PAGE,fusion protein polypeptide was extracted and purified,immunocompetence of the expressed multi-epitope protein was identified by Westernblot.BALB/c mouse was immuned by fusion protein polypeptide,the antibody was determined by ELISA,immunogenicity was evaluated.Results showed that expression recombinant plasmid pEGX-4T-1-ep contained with seven PCV2 antigen epitopes had been constructed successfully.SDS-PAGE analysis showed that fusion protein polypeptide was expressed effectively in Escherichia coli（E.coli）,and the molecular weight was about 35ku,which existed in the form of solubility.Results of Westernblot showed that the extraction and purification of fusion protein polypeptide and PCV2 positive serum had good reactogenicity.Results of ELISA showed that the purified fusion protein polypeptide could stimulate the body to produce PCV2 specific antibody which had good immunogenicity.Results indicated that the constructed PCV2 multi-epitope had good expression characteristics in vitro,and the expression protein had good immunogenicity.The study provided a basic for the study on PCV2 epitope screening,functional identification and multi-epitope vaccine.
Bibliography:In order to obtain induction expressed porcine circovirus type 2（PCV2）multi-epitope with good immunogenicity in vitro.Major epitopes of PCV2 were screened in the test,epitopes were composed sequentially,the c DNA sequences were artificially synthesized.Bam HⅠand SalⅠwere directionally cloned into prokaryotic expression vector PEGX-4T-1 multiple cloning site,then BL21 competent cells were transformed,positive clones were screened,IPTG inducible expression was conducted.Expression on target gene was analyzed by SDS-PAGE,fusion protein polypeptide was extracted and purified,immunocompetence of the expressed multi-epitope protein was identified by Westernblot.BALB/c mouse was immuned by fusion protein polypeptide,the antibody was determined by ELISA,immunogenicity was evaluated.Results showed that expression recombinant plasmid pEGX-4T-1-ep contained with seven PCV2 antigen epitopes had been constructed successfully.SDS-PAGE analysis showed that fusion protein polypeptide was expressed effectively in Escherichia coli（E.coli）,and the molecular weight was about 35ku,which existed in the form of solubility.Results of Westernblot showed that the extraction and purification of fusion protein polypeptide and PCV2 positive serum had good reactogenicity.Results of ELISA showed that the purified fusion protein polypeptide could stimulate the body to produce PCV2 specific antibody which had good immunogenicity.Results indicated that the constructed PCV2 multi-epitope had good expression characteristics in vitro,and the expression protein had good immunogenicity.The study provided a basic for the study on PCV2 epitope screening,functional identification and multi-epitope vaccine.
Porcine circovirus type 2 Multi-epitope Induced expression Immunogenicity
ISSN:1943-9911