猪繁殖与呼吸综合征病毒贵州株GZ-R NSP2基因的克隆与遗传变异分析

doi:10.16431/j.cnki.1671-7236.2020.11.029

Abstract

Abstract: In order to investigate the genetic variation of porcine reproductive and respiratory syndrome virus (PRRSV) strain GZ-R and the characteristics of NSP2 gene,in this study,two specific primers were designed based on NSP2 gene for nested RT-PCR,cloning and sequence analysis of the target gene.The results showed that the second round of nested PCR not only amplified the expected band of about 3 000 bp,but also amplified another band of 1 200 bp.The two DNA fragments were cloned into the pMD19-T vector,and two fragments were obtained by sequencing.The large fragment was 2 980 bp long and named as GZR-NSP2-L.The small fragment was 1 131 bp long and named as GZR-NSP2-S.The homology comparison results of the two fragments showed that GZR-NSP2-S had GZR-NSP2-L N-terminal nucleotide 1-945 and C-terminal 2 795-2 980 nucleotide,and lack of nucleotide between bits 946-2 794 in GZR-NSP2-L.The nucleotide homology of GZR-NSP2-L with VR-2332 strain and Lelystad virus strain was 81.2% and 48.6%.The results of homology comparison of encoded amino acids were consistent with the results of nucleotide homology,revealing that the PRRSV GZ-R strain belonged to the American-type strain.Comparison of amino acid deletion sites revealed that GZ-R NSP2 had 30 discontinuous amino acid deletions at positions 481 and 533-561.The deletion sites were consistent with PRRSV highly pathogenic strains.In summary,the PRRSV GZ-R strain could be identified as American-type highly pathogenic strain.The experimental results could provide some references for further exploring the epidemic trend of PRRSV in Guizhou province and the mechanism of NSP2 in PRRSV replication.

Key words: porcine reproductive and respiratory syndrome virus (PRRSV); NSP2; genetic variation analysis

CLC Number:

S855.3

ZHANG Xiyi, WEN Guilan, GUAN Guodan, CHEN Guang, TIAN Lang, YANG Baiqi. Cloning and Genetic Variation Analysis of NSP2 Gene of Porcine Reproductive and Respiratory Syndrome Virus Guizhou Strain GZ-R[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(11): 3659-3666.

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Cloning and Genetic Variation Analysis of NSP2 Gene of Porcine Reproductive and Respiratory Syndrome Virus Guizhou Strain GZ-R

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