摘要
[目的]研究四川资阳脑膜炎病例猪链球菌2型分离株ZYH33溶血素（suliysin,sly）基因的克隆表达及融合蛋白生物活性分析。[方法]从sly中获取第230~593氨基酸残基区域的基因片段,克隆至pMD18-T载体并鉴定,以重组pMD18-T/sly为模板PCR扩增基因片段,与表达载体pQE-30连接,转化至大肠杆菌TG1,重组子经PCR、酶切、测序鉴定。IPTG诱导重组蛋白表达,Western blot法鉴定融合蛋白的抗原性。[结果]基因片段在大肠杆菌中得到了表达,表达的融合蛋白可被猪链球菌2型菌体ZY05719抗血清识别,具有抗原性。[结论]所克隆、表达的溶血素区域可作为猪链球菌的诊断抗原,为基因工程疫苗的研制奠定了基础。
[Objective] To study human Streptococcus suis serotype 2 suilysin fusion protein and investigate its immunological activity.[Method] sly gene was obtained from ZYH33,which was isolated from human meningitis in Ziyang of Sichuan province.The gene was amplified by PCR with the template pMD18-T/sly.The gene fragments and expressing vector pQE-30 were ligated.The recombinant vector were transformed into TG1.The correct recombinations were identified by PCR,enzyme digestion and sequencing.Recombination protein and the peptides were detected by SDS-PAGE after induced by IPTG.The antigenicity of the recombination protein was determined with Western blot indicated by rabbit anti-ZY05719 strain polyclonal antibody.[Result]The sly gene fragments could expressed and the products were recognized by rabbit-ati Streptococcus suis serotype 2 polyclonal antibody.[Conclusion]This work demonstrated that the fragment encoding 230 to 593 amino acid resides of suilysin could be an effective antigen candidate for Streptococcus suis diagnosis and may be useful in the development of diagnostic reagents and genetic engineering vaccines.
出处
《安徽农业科学》
CAS
北大核心
2011年第6期3604-3605,3614,共3页
Journal of Anhui Agricultural Sciences
基金
山东省教育厅科技计划项目(J08LF57)
2010年度高等学校优秀青年教师国内访问学者项目
山东省聊城市科技攻关计划项目(20082048)资助
关键词
猪链球菌2型
溶血素
克隆
表达
抗原性
Streptococcus suis serotype 2
Suilysin
Clone
Expression
Antigenicity
