Prokaryotic Expression and Immunoreactivity Identification of Different Encoding Regions of the ORF5 Gene of PRRSV

doi:10.3321/j.issn:1000-7091.2007.03.004

Abstract

Abstract: To identify prokaryotic expression of the ORF5 gene of porcine reproductive and respiratory syndrome virus (PRRSV) in different encoding regions, fragments D, L, N and C obtained by deleting the N-terminal signal peptide sequence and/or the transmembrane domain sequence of ORF5 gene were amplified by PCR. Based on cloning and sequence. These fragments were inserted into the prokaryotic expression vector pET-32a respectively. The recombinant expression plasmids contained fragments L, N or C yielded recombinant fusion proteins of the expected sizes (33, 25 and 29 kDa respectively), but fragment D yielded no recombinant fusion protein. The recombinant fusion protein of fragment L showed strong positive reaction with PRRSV-infected swine serum by Western blotting, fragment C showed weak positive, whereas fragment N showed no positive. It indicated that the C-terminus of ORF5 encoding protein played an important role in immunoreactivity.

Key words: PRRSV, ORF5 gene, Prokaryotic expression, Immunoreactivity, Transmembrane domain

CLC Number:

Q786

ZHOU Hong-zhuan, XU Fu-zhou, WANG Jin-luo, YANG Xin-jian, SHI Ai-hua, YANG Bing. Prokaryotic Expression and Immunoreactivity Identification of Different Encoding Regions of the ORF5 Gene of PRRSV[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2007, 22(3): 12-15. doi: 10.3321/j.issn:1000-7091.2007.03.004.

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