红斑丹毒丝菌C43065株spaA基因的克隆和表达

doi:10.3969/j.issn.1007-2985.2013.05.021

journal6 ›› 2013, Vol. 34 ›› Issue (5): 85-88.DOI: 10.3969/j.issn.1007-2985.2013.05.021

红斑丹毒丝菌C43065株spaA基因的克隆和表达

刘丹丹, 杨振龙, 吾鲁木汗?那孜尔别克

（吉首大学生物资源与环境科学学院，湖南吉首 416000）

出版日期:2013-09-25 发布日期:2013-11-04
通讯作者: 吾鲁木汗·那孜尔别克，吉首大学生物资源与环境科学学院教授，博士，从事畜禽传染病免疫预防，E-mail：ulum@jsu.edu.cn.
作者简介:刘丹丹（1987-），女，湖南邵东人，吉首大学生物资源与环境科学学院硕士研究生，主要从事微生物生态学研究
基金资助:
国家自然科学基金资助项目（31072142）

Cloning and Expression of spaA Gene of Erysipelothrix Rhusiopathiae C43065

LIU Dan-Dan, YANG Zhen-Long, WU Lu-Mu-Han-·Na-Zi-尔Bie-Ke

(College of Biology and Environmental Sciences,Jishou University,Jishou 416000,China)

Online:2013-09-25 Published:2013-11-04

摘要/Abstract

摘要：通过PCR从红斑丹毒丝菌C43065株基因组DNA中扩增出编码信号肽除外的成熟SpaA蛋白基因spaA，将其克隆到表达载体pET32a的BamHⅠ和Hind Ⅲ位点上，构建重组表达质粒pET-spaA，转化大肠杆菌BL21，在IPTG诱导下表达N端带有Trx标签的融合蛋白rSpaA，SDS-PAGE检测表达蛋白.DNA测序结果表明，spaA基因大小为1794 bp，编码由597个氨基酸残基组成的成熟SpaA蛋白，SDS-PAGE结果显示在大肠杆菌BL21中成功表达了分子量约为86 kDa的重组rSpaA，为进一步开展SpaA保护区域的研究奠定基础.

关键词： 红斑丹毒丝菌, spaA基因, 克隆, 原核表达

Abstract: The spaA gene encoding mature surface protective antigen A (SpaA) without signal peptide was amplified from genomic DNA of E.rhusiopathiae C43065 by PCR,The BamHⅠand HindⅢ digested PCR product was cloned into prokaryotic expression vector pET32a to generate a recombinant plasmid pET-spaA.The recombinant protein rSpaA was expressed in E.coli BL21 harboring the recombinant plasmid pET-spaA by IPTG inducing,and the expressed protein was determined by SDS-PAGE.The DNA sequence analysis showed that the spaA gene of C43065 strain was 1794 bp in length.SDS-PAGE analysis revealed a single protein band with a molecular weight of 86 kDa successfully expressed in E.coli BL21.The expressed protein of rSpaA will contribute to further study on protective domain of this protein.

Key words: Erysipelothrix rhusiopathiae, spaA gene, cloning, prokaryotic expression

刘丹丹, 杨振龙, 吾鲁木汗?那孜尔别克. 红斑丹毒丝菌C43065株spaA基因的克隆和表达[J]. journal6, 2013, 34(5): 85-88.

LIU Dan-Dan, YANG Zhen-Long, WU Lu-Mu-Han-·Na-Zi-尔Bie-Ke. Cloning and Expression of spaA Gene of Erysipelothrix Rhusiopathiae C43065[J]. journal6, 2013, 34(5): 85-88.

参考文献

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[5] MAKINO S,YAMAMOTO K,MURAKAMI S,et al.Properties of Repeat Domain Found in a Novel Protective Antigen,SpaA,of Erysipelothrix Rhusiopathiae [J].Microb. Pathog,1998,25(2):101-109.

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[7] 吾鲁木汗·那孜尔别克,张磊,何翠,等.猪丹毒丝菌天然SpaA和重组SpaA-N免疫保护效果的评价 [J].微生物学报,2010,50(3):367-372.

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[10] 吾鲁木汗·那孜尔别克,刘祝祥,李科,等.猪丹毒丝菌C43311株spaA基因N端免疫保护区的克隆和表达 [J].微生物学报，2008,48(2):207-212.

红斑丹毒丝菌C43065株spaA基因的克隆和表达

Cloning and Expression of spaA Gene of Erysipelothrix Rhusiopathiae C43065

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