鹅细小病毒河北流行株HBZF07 NS1基因的克隆与序列分析

鹅细小病毒HBZF07株经13日龄鹅胚增殖后收集尿囊液,提取病毒基因组总DNA,采用PCR方法,一次性扩增出与预期大小相符的特异性条带.将扩增产物提纯回收后克隆入pMD18-T载体,经转化、筛选及酶切鉴定后,对阳性克隆进行了序列测定和序列分析.测序后拼接的基因长1 892 bp,包含完整的NS1开放阅读框(1 884 bp),编码623个氨基酸.与GenBank中其它毒株的NS1基因核苷酸序列相比,同源性分别为DY(EF515837)98.4%,与B(GPU25749)为99.2%,与HG5/82(AY506546)为93.9%,与YG(AF416726)为93.9%,与SHM319(GPU34761)为97.0%.

Cloning and Sequence Analysis of the NS1 Gene of Goose Parvovirus Hebei Isolate HBZF07

The Goose Parvovirus isolate HBZF07 was propagated in 13-day-old geese embryos. The allantonic fluid of the virus were concentrated and the genomic DNA was extracted. The NS1 gene of the virus was successfully amplified by PCR,and further cloned into pMD18-T vector,and a positive recombitant plasmid was screened by restriction enzyme analysis. The sequence analysis showed that the nucleotide sequence of this F gene was 1 884 bp and encoding a protein of 623 amino acids. Sequence comparision with other GPV strains selected from GenBank revealed that the HBZF07 NS1 gene had a high sequence homology to those of other GPV strains,98.4% with DY(EF515837),99.2% with B(GPU25749),93.9% with HG5/82(AY506546),93.9% with YG(AF416726),and 97.0% with SHM319(GPU34761).