一株新城疫病毒HN基因的克隆及序列分析

摘要

The HN gene of Newcastle disease virus isolate (NDV) JZ05 was amplified with reverse transcription-polymerase chain reaction (RT-PCR) , its gene cloning was completed, and its nucleotide sequence was determined. Alignment was executed to the HN gene and its amino acids sequence between NDV isolate JZ05 and other 18 NDV isolates by a sequence analysis software ( namely DNAssist). The result of sequence determining to the HN gene showed that there were 1716 bp in the HN gene, and in HN protein there were 571 amino acid residues, 5 sites of potential carbohydrate binding, 13 cysteine residues, no sixth site (538-540 aa). In HN protein of NDV isolate JZ05 single amino acid residue of 6 sites became different as compared with other 18 NDV isolates. Homology was 94% over for the HN gene nucleotide sequence and amino acid sequence between strain JZ05 and 11 strains (ZJ1, TJ03, Jlan-1-00, SBZ02, SCL03, JS02, GD05, SQD04, SQZ04, YG, Taiwan 95) , but homology was about 82% for HN gene nucleotide sequence and about 84% for its amino acid sequence between strains JZ05 and 3 vaccine strains (Clone30, LaSota, B1).%以RT-PCR扩增新城疫病毒JZ05株的HN基因,并进行克隆和测序.采用DNAssist序列分析软件将JZ05毒株HN基因与另外18个NDV毒株HN基因序列及其氨基酸序列进行比对分析.测序结果表明,JZ05毒株的HN基因全长1716 bp,HN蛋白由571个氨基酸残基构成,具有5个潜在糖基化位点及13个半胱氨酸残基,第6个潜在糖基化位点(538-540 aa)发生缺失.与另外18个NDV毒株比对,仅NDV JZ05株发生单个氨基酸残基变更的有6处.与11个毒株(ZJ1、TJ03、Jlan-1-00、SBZ02、SCL03、JS02、GD05、SQD04、SQZ04、YG、Taiwan95)比对,HN基因序列及其氨基酸序列的同源性均在94%以上;但与3个疫苗毒株(Clone30、LaSota、B1)比对,基因序列的同源性约82%,氨基酸序列的同源性约87%.