摘要
为克隆、表达犬链球菌(S.canis)保护性抗原基因,并对其表达产物进行免疫原性分析。本研究以Lambda ZAPⅡ载体构建了S.canis的3kb～8kb随机基因组文库,通过S.canis阳性血清筛选和质粒拯救,获得一株阳性克隆重组质粒并进行了序列测定。通过BLAST分析并与GenBank中登录的相近基因序列比较发现,该基因为S.canis的1个保护性抗原(SPASc)基因,该基因与兽疫链球菌和产脓链球菌的保护性抗原及马链球菌M蛋白基因具有一定的同源性。根据该序列设计特异性引物,经PCR扩增、克隆并进行了SPASc基因的原核表达。用纯化SPASc重组蛋白免疫兔制备的血清对小鼠具有保护性作用。
Streptococcus canis （S.canis） is an important opportunistic pathogen that causes infections among a variety of animals including human. A new protective antigen（SPASc） was identified by screening a S.canis phage library of 3 kb-8 kb random DNA fragments. The sequence of the SPASc was determined by sequencing the rescued plasmids. The SPASc gone showed homology to that of Streptococcus equi subsp, zooepidemicus, Streptococcus pyogenes, and M protein of Streptococcus equi. Rabbit antiserum to recombinant SPASc was shown to protect mouse from S.canis challenge.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2009年第9期680-683,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
天津市自然科学基金项目(07JCYBJC16000)
