鹅细小病毒VP1蛋白氨基酸点突变在鹅胚化疫苗株毒力减弱中的关键作用

发布时间：2018-02-11 03:56

本文关键词： 鹅细小病毒疫苗株点突变嵌合病毒毒力　出处：《中国兽医学报》2016年12期 　论文类型：期刊论文

【摘要】：为了进一步明确各编码蛋白基因在鹅细小病毒(goose parvovirus)鹅胚化疫苗株毒力减弱中的作用,以先前构建的弱毒疫苗株感染性质粒pSYG61v和强毒株感染性质粒pLH为基础,通过重叠PCR方法,在pSYG61v和pLH质粒之间互换Rep1和VP1基因片段,获得了6个嵌合质粒pSYG-vRep1、pSYG-vVP1、pSYG-vRC、pLH-aRep1、pLH-aVP1和pLH-aRC。将质粒以绒毛尿囊膜途径转染11日龄鹅胚分别拯救出嵌合病毒。嵌合病毒的雏鹅攻毒试验表明,携带强毒株VP1基因的pSYG-vRC、pSYG-vVP1和pLH-aRep1这3个嵌合病毒对雏鹅表现出明显致病性,死亡率在75.0%~87.5%。完全携带弱毒株rep-cap编码框的pLH-aRC嵌合病毒攻毒组雏鹅无一死亡。携带强毒株Rep1基因的pLH-aVP1和pSYG-vRep1嵌合病毒攻毒组雏鹅也无一死亡,但在试验期内部分雏鹅明显生长迟缓。嵌合病毒攻毒试验结果表明,结构蛋白VP1基因的氨基酸点突变对GPV鹅胚化疫苗株的毒力减弱起着关键作用。
[Abstract]:In order to further clarify the role of various coding protein genes in the reduction of virulence of goose embryo vaccine strain of goose parvovirus, based on the previously constructed attenuated vaccine strain pSYG61v and virulence plasmid pLH of virulent strain, overlapping PCR method was used. Six chimeric plasmids pSYG-vRep1pSYG-vVP1npSYG-vRC+ pLH-aVP1 and pLH-aRCwere obtained by exchanging Rep1 and VP1 gene fragments between pSYG61v and pLH plasmids. Chorioallantoic membrane was used to transfect the plasmids into 11 day-old goose embryos to save chimeric viruses. Three chimeric viruses, pSYG-vRCU pSYG-vVP1 and pLH-aRep1, which carry the VP1 gene of virulent strain, showed obvious pathogenicity to goslings. The mortality rate was 75.0% and 87.5%. None of the goslings in the pLH-aRC chimeric virus attack group carrying the attenuated strain rep-cap coding frame had died, and none of the goslings in the pLH-aVP1 and pSYG-vRep1 chimeric virus groups carrying the Rep1 gene of the virulent strain had died. The results showed that the amino acid point mutation of structural protein VP1 gene played a key role in weakening the virulence of GPV goose embryo vaccine strain.
【作者单位】：扬州大学兽医学院江苏省动物重要疫病与人兽共患病防控协同创新中心;
【基金】：国家自然科学基金资助项目(31172317;31572511) 江苏省高校优势学科建设工程资助项目
【分类号】：S852.65

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