Constructing gE, gI and US9 gene deletion strain of pseudorabies virus

doi:10.3969/j.issn.1004-1524.2017.04.04

›› 2017, Vol. 29 ›› Issue (4): 542-547.DOI: 10.3969/j.issn.1004-1524.2017.04.04

• Animal Science • Previous Articles Next Articles

Constructing gE, gI and US9 gene deletion strain of pseudorabies virus

FAN Yi¹, LI Bi¹, GUO Wanzhu¹, LI Ping¹, HUANG Jianbo¹, YANG Fan¹, JIANG Ziyi¹, ZHAO Jun¹, XU Siyao¹, DENG Yichao¹, YIN Yue¹, MAO Xiyu¹, LYU Wenting¹, XU Zhiwen^{1, 2}, ZHU Ling^{1, 2, *}

1. Veterinary Medicine College of Sichuan Agricultural University, Chengdu 611130, China;
2. Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu 611130, China

Received:2016-11-13 Online:2017-04-20 Published:2017-04-27

Abstract

Abstract: Pseudorabies virus (PRV), a member of Herpes virus has an extremely broad range in nature and threatens the pig-industry development in our country. To explore the mechanism of nerve conduction of PRV, a virus mutant with a deletion in gE, gI and US9 genes was constructed. Plasmid of PP63 and pseudorabies virus Fa strain genomic DNA were co-transfected into BHK-21 cells by using Lipofectamine 3000 transfection reagent, and a recombinant virus with the deletion of gI/gE/US9 , named SA215-T, was screened and purified by plaque assay. PCR, gene sequencing. Western Blot, electron microscope and growth curve were used for identification of the deletion of genes, and features of the recombinant. The results showed that SA215-T was with effective deletion of gI and gE gene by PCR,and was absent in the expression of gE gene by Western Blot. There were no obvious differences in the morphology and growth curve of the recombinant virus as compared to its parental virus, they both achieved a high viral titer in cells.

Key words: pseudorabies virus, gI/gE/US9 gene, recombinant virus, gene deletion

CLC Number:

S852.65

FAN Yi, LI Bi, GUO Wanzhu, LI Ping, HUANG Jianbo, YANG Fan, JIANG Ziyi, ZHAO Jun, XU Siyao, DENG Yichao, YIN Yue, MAO Xiyu, LYU Wenting, XU Zhiwen, ZHU Ling. Constructing gE, gI and US9 gene deletion strain of pseudorabies virus[J]. , 2017, 29(4): 542-547.

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Constructing gE, gI and US9 gene deletion strain of pseudorabies virus

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