摘要
参考GenBank收录的马动脉炎病毒(EAV)读码框大囊膜蛋白基因 ORF5、膜蛋白基因ORF6的核苷酸序列,分别设计了2对引物,对EAV的这两种主要结构蛋白基因进行了 RT PCR;将扩增产物克隆于pUC18通用载体,对重组质粒进行限制性内切酶分析和基因测序,证实克隆片段的可靠和准确性。测序结果,ORF5目的片段包含768 bp,编码255个氨基酸组成的多肽;ORF6目的片段包含489 bp,编码162个氨基酸组成的多肽。与EAV NC 002532标准毒株比较,ORF5、ORF6核苷酸的同源性分别为99.1%和99.4%;推导氨基酸的同源性分别为96.9%和98.2%。
Two pairs of primers for RT-PCR were designed according to the sequences from GenBank to amplify major envelope protein gene and membrane protein gene of equine arteritis virus.The amplicons were cloned subsequently into the pUC18 plasmid. The recombinant plasmids pUC18-GL and pUC18-M were identified by restriction enzyme digestion and sequencing. Nucleotide sequences analysis showed that the major envelope protein gene contains an open reading frame of 768bp, encoding a 255 aa protein and the membrane protein contains an open reading frame of 489bp, encoding a 162 aa protein,respectively. The comparison of the two genes with that of EAV NC-002532 strain showed that homology of nucleotide sequence were 99.1% and 99.4%, respectively; and homology of deduced amino acids were 96.9% and 98.2%, respectively.
出处
《中国兽医科技》
CSCD
北大核心
2005年第2期112-116,共5页
Chinese Journal of Veterinary Science and Technology
