Construction of recombinant PRRSV ORF5 gene by DNA shuffling technique.

Author: Jiankui LIU ¹ ; Chunhua WEI ¹ ; Chunfang HUANG ¹ ; Xiaoyan CHEN ¹ ; Ailing DAI ¹ ; Xiaoyan YANG ¹
Author Information

1. Fujian Engineering Research Center for Swine Disease Control and Prevention, Fujian Provincial Key Laboratory for the Prevention and Control of Animal Infectious Diseases and Biotechnology, College of Life Sciences of Longyan University, Longyan 364000, Fujian, China.
Publication Type:Journal Article
Keywords: DNA shuffling; ORF5 gene; PRRSV; inhibition assay of virus infection; polyclonal antibody
From: Chinese Journal of Biotechnology 2018;34(6):888-896
CountryChina
Language:Chinese
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.