| 猪链球菌2型分解代谢控制蛋白A的克隆和原核表达 |
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| 引用本文： | 冉会玲,郎需龙,王兴龙,张瑞,钱晶,陈思,王秀然.猪链球菌2型分解代谢控制蛋白A的克隆和原核表达[J].中国畜牧兽医,2013,40(3):7-10. |
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| 作者姓名： | 冉会玲 郎需龙 王兴龙 张瑞 钱晶 陈思 王秀然 |
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| 作者单位： | 1. 吉林农业大学动物科技学院,吉林长春 130118;2. 军事医学科学院军事兽医研究所,吉林长春 130122;3. 吉林省人兽共患病预防与控制重点实验室,吉林长春 130122 |
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| 基金项目： | 国家自然科学基金项目(31101790/C1803)。 |
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| 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。
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| 关 键 词： | 猪链球菌2型 ccpA基因 原核表达 |
| 收稿时间： | 2012-09-27 |
Cloning and Prokaryotic Expression of ccpA gene of Streptococcus suis Type 2 |
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| RAN Hui-ling,LANG Xu-long,WANG Xing-long,ZHANG Rui,QIAN Jing,CHEN Si,WANG Xiu-ran.Cloning and Prokaryotic Expression of ccpA gene of Streptococcus suis Type 2[J].China Animal Husbandry & Veterinary Medicine,2013,40(3):7-10. |
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| Authors: | RAN Hui-ling LANG Xu-long WANG Xing-long ZHANG Rui QIAN Jing CHEN Si WANG Xiu-ran |
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| Institution: | 1. College of Animal Science and Technology, Jilin Agricultural University, Changchun 130118, China;2. Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun 130122, China;3. Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Changchun 130122, China |
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| Abstract: | 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. |
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| Keywords: | Streptococcus suis type 2 ccpA gene prokaryotic expression |
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