摘要
为了研究吉林省猪圆环病毒3型(PCV3)野毒株遗传变异情况及其衣壳蛋白(Cap)的亚细胞定位特征,本试验对收集到的10株PCV3吉林野毒株进行全基因组序列测定和分析,并构建真核表达载体pcDNA3.1V5/HisA-Cap转染猪肺巨噬细胞(PAM);继而用本实验室已构建的兔源PCV3-Cap多克隆抗体,通过激光共聚焦检测Cap蛋白在PAM中的定位。序列分析结果显示,10株PCV3吉林野毒株与国内外参考株之间的全基因组核苷酸序列同源性为95.8%~99.5%,处于PCV3a和PCV3b两个亚群;基因重组分析结果显示,仅中国参考株MF589102(2016年)、MH445396(2019年)和分离株JL-2在80~303 nt、840~1196 nt和1885~2001 nt处发生3处重组。激光共聚焦检测结果显示,Cap蛋白大部分定位于细胞核,少部分定位于细胞质。结果表明,目前吉林省PCV3比较保守,没有出现较大变异,Cap蛋白在PAM的细胞核和细胞质中均有分布。
In order to determine the genetic variation of Jilin porcine circovirus type 3(PCV3)wild strains and the subcellular localization of its capsid(Cap)protein,this study analyzed the whole gene sequences of 10 Jilin PCV3 wild strains collected in the laboratory,constructed the eukaryotic expression vector pcDNA3.1V5/HisA-Cap for transfection into porcine pulmonary macrophages(PAM),and applied rabbit anti-PCV3 Cap polyclonal antibody prepared previously in the laboratory for laser confocal localization of Cap protein in cells.The sequence analysis results showed that the nucleotide homology of the whole gene sequences between 10 Jilin PCV3 wild strains and domestic and foreign reference strains was 95.8%-99.5%,and the PCV3 strains examined in this study were of subgroups PCV3a and PCV3b.The gene recombination results showed that the JL-2,the 2016 Chinese reference strain MF589102 and the 2019 Chinese reference strain MH445396 had three recombinations at 80-303 nt,840-1196 nt and 1885-2001 nt.Laser confocal results showed that most of the Cap proteins were located in the nucleus and a few in the cytoplasm.These results indicate that PCV3 is relatively conservative in relation to other viruses with limited variation,Cap protein exists mainly in the nucleus and occasionally in the cytoplasm.
作者
刘东旭
刘宏莹
苏诺
葛桂阳
麻宝艺
盛陈艳
李东丽
刘艺
LIU Dong-xu;LIU Hong-ying;SU Nuo;GE Gui-yang;MA Bao-yi;SHENG Chen-yan;LI Dong-li;LIU Yi(Jilin Agricultural Science and Technology College,Jilin 132101,China;College of Animal Science and Technology,Jilin Agricultural University,Changchun 130118,China)
出处
《中国兽医杂志》
CAS
北大核心
2023年第4期27-32,共6页
Chinese Journal of Veterinary Medicine
基金
吉林省教育厅“十三五”科学技术项目(JJKH20200394KJ)
吉林农业科技学院青年基金项目[吉农院合字(20200002)号]。
关键词
野毒株
猪圆环病毒3型
CAP蛋白
亚细胞定位
wild strain
porcine circovirus type 3
Cap protein
subcellular localization
