摘要
原核表达猪链球菌2型分解代谢控制蛋白A(catabolite control protein A,ccpA)并进行纯化,通过免疫印迹反应初步鉴定重组蛋白的抗原性和特异性。采用PCR方法,扩增ccpA基因序列,克隆至原核表达载体pET-28a中,转入大肠杆菌BL21中进行诱导、表达和亲和层析纯化;经Western blotting初步评价ccpA的抗原性。重组原核表达质粒pET-28a-ccpA经双酶切及测序证明构建正确;表达的重组蛋白相对分子质量约38000,能被相关抗体所识别。因此,能成功在大肠杆菌中表达猪链球菌2型ccpA,为猪链球菌2型相关调节蛋白的免疫学研究奠定基础。
To obtain recombinant ccpA of Streptococcus suis type 2 expressed in prokaryotic system and preliminaryly evaluate its antigenicity and specificity by Western blotting.ccpA gene was obtained by PCR and cloned into pET-28a vector,then transformed into Escherichia coli BL21 to induce,express and purify,the antigenicity of recombinant ccpA was identified by Western blotting.The double enzyme analysis and sequencing proved that recombinant plasmid pET-28a-ccpA was constructed correctly.The expressed recombinant protein,with a relative molecular mass of about 38000,was recognized by positive serum antibody.The ccpA of Streptococcus suis type 2 was successfully expressed in E.coli,which provided a material for development of serological diagnostic method for Streptococcus suis type 2.
出处
《中国畜牧兽医》
CAS
北大核心
2013年第3期7-10,共4页
China Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金项目(31101790/C1803)
关键词
猪链球菌2型
ccpA基因
原核表达
Streptococcus suis type 2
ccpA gene
prokaryotic expression
