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| Preparation of Polyclonal Antiserums of GP5a Protein of Porcine Reproductive and Respiratory Syndrome Virus |
| WANG Qing, XU Yan-zhao, WEI Xiao-xiao, WANG Qiu-xia, HANG Bo-lin, SUN Ya-wei, WANG Fei-fei, HU Jian-he |
| Henan Institute of Science and Technology, Xinxiang 453003, China |
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Abstract According to the sequences of GP5a gene of XX2012 strain HP-PRRSV, a pair of specificity primers were designed synthesis. The complete gene fragment of GP5a protein was amplified by RT-PCR, and the eukaryotic expression vector pCAGGS-GP5a which containing GP5a protein gene was constructed. Polyclonal antiserums of GP5a protein were prepared by gene immunization pCAGGS-GP5a vector in BALB/c mice. The results showed that the sequences of GP5a protein gene were consistent with predicted size, 156 base pairs. PCR, digestion and sequencing results showed that the recombinant plasmid pCAGGS-GP5a was correct. After the mice were immuned with pCAGGS-GP5a three times, the separated serums of these mice contain the polyclonal antibody of GP5a protein, and the results of indirect immuno-fluorescence assay (IFA) showed that the polyclonal antiserums can specifically recognize cells infected by the virus. In summary, the recombinant plasmid pCAGGS-GP5a containing GP5a protein gene was constructed, and the polyclonal antibody of GP5a protein was prepared, which laid the foundation for the further study of subcellular localization and relevant function of GP5a protein.
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Received: 06 March 2015
Published: 25 August 2015
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