1. [Construction of recombinant PRRSV ORF5 gene by DNA shuffling technique].
- Author
-
Liu J, Wei C, Huang C, Chen X, Dai A, and Yang X
- Subjects
- Animals, Antibodies, Viral, Escherichia coli, Mice, Mice, Inbred BALB C, Recombinant Proteins genetics, DNA Shuffling, Porcine respiratory and reproductive syndrome virus genetics, Viral Envelope Proteins genetics
- Abstract
Recombinant PRRSV △2ORF5 gene was constructed using DNA shuffling from four genetically different strains of PRRSV to study its heterologous cross-neutralizing ability. The △2ORF5 mutant gene was cloned into the vector pET-32a and transferred into E. coli BL21. SDS-PAGE confirmed that the molecular weight of the recombinant △2ORF5 was about 42 kDa, consistent with the predicted result. Then the purified recombinant protein was injected into BALB/c mouse to obtain polyclonal antibody. Western blotting analysis with mouse-anti-△2ORF5 polyclonal serum indicated that the parental virus recombinant GP5 protein reacted with the specific antibodies. Four parental viruses could be inhibited by the anti-△2ORF5 polyclonal antibody and the inhibition rates were higher than 53%. This work has laid a foundation for further development vaccine for PRRSV.
- Published
- 2018
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