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。
Abstract:
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.
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