摘要
为进一步认识AEV的基因结构及功能,为基因工程苗的研制,以及分子流行病学研究提供理论基础,本研究根据已发表的AEV序列设计一对引物,利用反转录———聚合酶链式反应(RT-PCR)技术,在体外从AEVHN株感染的SPF鸡胚病变组织中扩增出VPO基因,经纯化后将VPO基因克隆到质粒pGEM-T-Easy上,构建成含有VPO基因的重组质粒pGEM-T-VPO,然后通过转化感受态大肠杆菌,经蓝白斑筛选,挑出阳性克隆,提取质粒做PCR和酶切鉴定。进一步序列分析表明,该基因长726bp,编码242个氨基酸,与AEVBZ株和HB株的核苷酸和氨基酸同源性分别为:99.2%、98.3%和94.8%、95.4%,同源性较高,说明VPO基因较为保守。
A pair of primers for avian encephalo myelitis virus(AEV)isolate HN was designed according to published AEV genome.With use of reverse transcription -Polymerase Chain Reaction(RT -PCR),fragment VPO was in vitro am-plified from SPF chicken embryo tissue after inoculation of AEV isolate HN.The fragment VPO gene was inserted into plasmid PGEM-T -Easy to obtain the re combinant plasmid VPO -PGEM-T.VPO gene sequence was 242aa.By com-parison with other published gene of AEV,the nucleotide se-quence and deduced amino acid of VPO o f the strain were99.2?98.3and 94.8?95.4with the correspondent sequences of AEV strain BZ and HB Respectively.This demonstrated that the VPO Gene was conservative.The experiment offered theo-retical foundation to the studying o f structure and function of AEV VPO gene,molecular epidemy and e ngineered vaccine.
出处
《河南畜牧兽医》
2002年第9期6-7,共2页
Henan Journal of Animal Husbandry and Veterinary Medicine
