摘要
根据GenBank登录的鸡新城疫病毒NP基因序列设计引物,利用RT-PCR对F48E9强毒株NP基因进行扩增,将扩增产物经纯化后连入pMD18-T载体,通过PCR和酶切鉴定出阳性重组质粒,经测序后获得F48E9 NP基因序列。经分析发现该基因的ORF总长为1 470 bp,编码489 aa,NP蛋白有4个高度保守的Cys位点,分别位于第55、78、139、213位,且在401～440位和461～481位区域变异较大。将F48E9株与19株参考毒株的相应序列进行比较得到核苷酸序列同源性为87.9%～92.3%,氨基酸序列同源性较高,为92.2%～98.0%,遗传进化分析表明F48E9株相对独立,与参考毒株的遗传距离较远。
The nucleocapsid protein gene of velogenic, chicken-origin, Newcastle disease virus （NDV） F48E9 was amplified by RT - PCR based on a specific pair of primers designed according to the chicken-origin NDVs NP gene sequences published at GenBank. The purified RT - PCR products were cloned onto the pMD18 - T vectors, and then, the positive recombinants had been identified by PCR and enzyme digestion. The sequencing results indicated that the F48E9 NP gene had an opening read frame of 1470nt in length with a coded polypeptide of 489 amino acids. There were four highly conserved cysteines on the F48E9 strain nucleocapsid protein. The amino acids at the regions, aa 401-440 and aa 461-481, were found to vary more than the others. The sequence analysis indicated that the F48E9 strain had a nucleotide homology of 87.9% -92.3%, and the amino acid homology of 92.2% -98.0% with other NDVs. The F48E9 strain appeared to be evolutionarily independent, with only remote relationship with other NDVs.
出处
《福建农业学报》
CAS
2009年第5期385-389,共5页
Fujian Journal of Agricultural Sciences
基金
国家自然科学基金资助项目(30671564)
国家"973"子项目(2005CB523001)
