【摘要】 根据已发表的鹅细小病毒(GPV)核苷酸序列,设计并合成了1对引物,对吉林农业大学预防兽医学研究室分离鉴定的GPV四平株(SP)主要保护性抗原蛋白VP3基因进行扩增。所得到的PCR产物片段大小约1.6 kb,与预期片段大小相符。将该片段纯化后与T载体连接,转化到感受态大肠杆菌JM-109中进行克隆。对所提质粒进行快速鉴定、PCR鉴定以及限制性内切酶NheⅠ和BamHⅠ双酶切鉴定,筛选阳性重组质粒,对重组质粒测序并进行核苷酸序列分析。结果表明:GPV四平株VP3基因长1 605 nt,包含了完整编码GPV VP3蛋白阅读框,且与国内GPV分离株B,GD,HG5/82相应核苷酸序列和氨基酸序列的同源性较高,分别为96.2%,96.3%,96.4%和97.4%,98.7%,98.9%。据此认为,GPV四平株VP3基因可以作为构建具有普遍应用价值的基因工程疫苗的候选基因。更多还原

【Abstract】 A pair of primers were designed and synthesized according to the nucleotide sequence of goose parvovirus(GPV) B strain and VP3 gene was amplified from the DNA of goose parvovirus isolated in Siping district,Jilin province.The PCR amplified VP3 gene was cloned into pMD18-T vector.The recombinant plasmid was identified with restriction endonucleases NheⅠ and BamHⅠ analysis,PCR and sequencing.The result of sequence analysis showed that VP3 gene was composed of 1 605 bp and included a complete open reading frame which encoded a protein of 534 amino acids,and the nucleotide sequences of isolate SP strain shared homology of 96.2%,96.3%,96.4% with the sequence of the B,GD,HG5/82 strain of GPV respectively.And amino acid sequences of isolate SP strain shared homology of 97.4%,98.7%,98.9% with the sequence of the B,GD,HG5/82 strain of GPV respectively.According to the results,the VP3 gene of GPV SP isolate could be used as candidate gene to construct gene engineering vaccine utilized widely.更多还原