摘要： 采用RT-PCR方法自犬瘟热病毒Lederle株基因组RNA中扩增出H基因(1911 bp)和N基因(1707 bp)片段，序列测定和分析表明与ONderstepoort株、CDV3株等疫苗毒株的同源性在95%以上，而与野外分离株的同源性介于90%～95%；以获得的H，N全基因片段为模板，分别扩增H基因近3’端996 bp的基因片段以及N基因中间高度保守区562 bp的基因片段，经克隆测序后分别插入原核表达载体pET-32a中进行表达，结果显示H片段和N片段分别表达大小约为57 kD和37 kD的融合蛋白，WesterN blottiNg检测结果显示，表达蛋白均可与CDV抗血清发生阳性反应，提示原核表达产物具有与CDV抗血清结合的免疫反应性。

关键词: 犬瘟热病毒, H基因, N基因, 序列分析, 原核表达

Abstract: The haemagglutinin protein gene (H) and the nucleocapsid protein gene (N) were amplified from the genomic RNA of canine distemper virus Lederle strain by RT-PCR. The full length of the H fragment and the N fragment was 1911 bp and 1707 bp respectively. Sequence analysis showed that the H and N gene had an over 95% identity with vaccine strains such as Onderstepoort strain and CDV3 strain, while only 90%-95% with wild isolates. A 996 bp H fragment and a 562 bp N fragment were amplified and inserted into the prokaryotic expression vector pET-32a for induced expression. The size of the recombinant fusion proteins was 57 kD (H) and 37 kD (N) respectively. The recombinant fusion proteins showed positive reaction with anti-CDV canine serum by Western blotting.

Key words: Canine distemper virus, H gene, N gene, Sequence analysis, Prokaryotic expression

中图分类号: 

• S852.65+5