根据Genbank中的鹅细小病毒(GPV)B株全基因序列,设计合成一对引物,应用PCR技术扩增了GPV强毒株CHv的VP3基因片段,将扩增后的VP3基因重组到pMD18-T质粒载体上,并对插入片段进行序列测定,将测序结果及由该结果推导的氨基酸序列与国内外分离的GPV、MDPV、PPV和CPV等不同宿主的细小病毒的VP3进行比对分析。结果表明:中国四川分离的GPV CHv株VP3基因长1 605 bp,编码534个氨基酸,与国内外10株GPV的VP3基因进行比较,核苷酸同源性为93.4%~99.8%,氨基酸同源性为96.5%~99.3%,其变异较小,是GPV保持一个血清型的分子基础。与番鸭细小病毒的核苷酸和氨基酸同源性分别为79.6%和89.9%,而与其他种属的细小病毒同源性均在30%以下,表明它们与GPV CHv株亲缘关系较远。
Primers were designed according to the genome of GPV B strain in Genbank.The DNA encoding VP3 structure protein of GPV CHv strain was amplified by PCR.The VP3 gene was cloned into pMD18-T vector and then sequenced.The DNA and amino acid sequence were compared with other GPV,MDPV,PPV,CPV and BPV.The results showed that the DNA of VP3 was 1 605 nt and coding 534 amid acids,shared 93.4%~99.8% nucleotide sequences identity with those published in Genbank,and shared 96.5%~99.3% amino acid sequence identity,which was the reasons that there was only one serum type of GPV.GPV CHv VP3 and MDPV shared 79.6% nucleotide sequences identity and 89.9% amino acid sequence identity,and shared less 30% nucleotide sequences identity with other parvovirus.