稻瘟病菌MSP1的表达、纯化及生物信息学分析  

Expression,Purification and Bioinformatics Analysis of Magnaporthe oryzae MSP1

作　　者：赵秀平 李国瑞[1] 王双[1] 闫星伊 段强 张帅 风兰 韩雯毓 孙佳欣 陈永胜[1] ZHAO Xiu-ping;LI Guo-rui;WANG Shuang;YAN Xing-yi;DUAN Qiang;ZHANG Shuai;FENG Lan;HAN Wen-yu;SUN Jia-xin;CHEN Yong-sheng(School of Life Science and Food Science,Inner Mongolia University for Nationalities,Inner Mongolia Industrial Engineering Research Center of Universities for Castor,Inner Mongolia Key Laboratory of Castor Breeding,Inner Mongolia Collaborative Innovation Center for Castor Industry,Inner Mongolia Engineering Research Center of Industrial Technology Innovation of Castor,Inner Mongolia Tongliao 028043,China)

机构地区：[1]内蒙古民族大学生命科学与食品学院,内蒙古自治区高校蓖麻产业工程技术研究中心,内蒙古自治区蓖麻育种重点实验室,内蒙古自治区蓖麻产业协同创新中心,蓖麻产业技术创新内蒙古自治区工程研究中心,内蒙古通辽028043

出　　处：《西南农业学报》2021年第7期1433-1438,共6页Southwest China Journal of Agricultural Sciences

基　　金：内蒙古民族大学重点实验室开放基金项目(MDK2019020)。

摘　　要：【目的】稻瘟病菌MSP1是由MGG-05344基因编码的一种与稻瘟病菌的植物毒性相关的蛋白,本研究进一步探索MSP1的晶体结构、功能以及与该蛋白相关的研究。【方法】对NCBI数据库提供的MSP1氨基酸序列进行生物信息学分析,构建原核表达载体pETSUMO_1b-MSP1,并对该重组蛋白进行诱导表达和纯化处理。【结果】MSP1分子量约为14.2 KD,等电点为4.6,为不稳定性疏水蛋白;含有信号肽,无跨膜结构域,为非典型分泌蛋白;含有11个磷酸活性位点,含有Cerato-platanin保守结构域,属于Cerato-platanin家族成员。该蛋白可被0.3 mmol/L IPTG诱导表达,镍离子亲和层析的最适洗脱条件为20 mmol/L Tris-HCl,500 mmol/L NaCl,80 mmol/L、100 mmol/L咪唑;阴离子交换层析表明该蛋白具有耐低盐性;分子筛层析具有单峰且对称性良好,最大峰在17.1 mL出峰,分子量约20 KD,表明MSP1以单体形式存在。【结论】本研究最终纯化得到大量高纯蛋白,以期为进一步探索该蛋白的晶体结构、功能、互作蛋白以及与稻瘟病菌毒性相关的后续研究奠定理论与实践基础。【Objective】Magnaporthe oryzae MSP1 is a protein encoded by MGG-05344 gene related to phytotoxicity of Magnaporthe oryzae,in order to further explore the crystal structure and function of MSP1 and the research related to this protein.【Method】In the present study,the amino acid sequence of MSP1 provided by NCBI database was analyzed by bioinformatics,the prokaryotic expression vector pETSUMO1 b-MSP1 was constructed,and the recombinant protein was induced,expressed and purified.【Result】The molecular weight of MSP1 was 14.2 KD,and its isoelectric point was 4.6.MSP1 was an unstable hydrophobin.It contained signal peptide,had no transmembrane domain and was an atypical secretory protein.It contained 11 phosphate active sites and cerato-platinin conserved domain,belonging to cerato-platinin family.The expression of the protein could be induced by 0.3 mmol/L IPTG,and the optimum elution conditions of nickel ion affinity chromatography were 20 mmol/L Tris-HCl,500 mmol/L NaCl and 80 mmol/L,100 mmol/L imidazole.The anion exchange chromatography showed that the protein had low salt tolerance.The molecular sieve chromatography had a single peak with good symmol/Letry,with the maximum peak at 17.1 mL and the molecular weight of about 20 KD,indicating that MSP1 existed as monomer.【Conclusion】A large number of high-purity proteins were purified in this study,in order to lay a theoretical and practical foundation for further exploring the crystal structure,function,interaction protein and subsequent research related to the toxicity of Magnaporthe oryzae.

关 键 词：稻瘟病菌 MSP1 MGG-05344 纯化 生物信息学分析 

分 类 号：S344.111.41[农业科学—作物栽培与耕作技术]