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2021, 04, v.40 1590-1596
稻瘟病菌MGG_14095基因克隆、表达及其表达产物的纯化
基金项目(Foundation): 内蒙古民族大学科研项目(MDK2016008; MDK2018018; MDK2019020); 内蒙古自治区高等学校科学研究项目(NJZY20121)共同资助
邮箱(Email): 15849551543@163.com;
DOI: 10.13417/j.gab.040.001590
摘要:

稻瘟病菌(Magnaporthe oryzae)是世界上危害性最大的水稻病原菌,也是阐明植物真菌病分子基础的主要模式生物,开展稻瘟病菌生长发育的相关研究,对稻瘟病的防治有重要意义。MGG_14095基因是稻瘟病菌的致病基因,对MGG_14095蛋白进行生物信息学预测,结果显示,MGG_14095蛋白等电点(pI)为5.72,不稳定指数为50.99,属于酸性不稳定蛋白;含角质酶保守结构域,隶属α/β水解酶超家族;有明显跨膜结构和信号肽剪切位点,属于分泌型蛋白质。本研究克隆稻瘟病菌MGG_14095(38-281)基因,构建原核表达系统pETM20-MGG_14095(38-281),采用分步层析纯化MGG_14095(38-281)蛋白,成功获得高纯度可溶目标蛋白。本研究结果为解析MGG_14095(38-281)蛋白质结构、探索稻瘟病菌致病机理及对稻瘟病菌防治提供一定的理论与试验依据。

Abstract:

Magnaporthe oryzae is the most harmful pathogen in the world for rice and the main model organism for elucidating the molecular basis of plant fungal disease. Carrying out relevant research of the growth and development of Magnaporthe oryzae has a great significance for the control of Magnaporthe oryzae. MGG_14095 gene is the pathogenic gene of Magnaporthe oryzae. The bioinformatics prediction of MGG_14095 protein showed that the isoelectric point(pI) of MGG_14095 protein was 5.72, and the instability index was 50.99, belonging to the acid unstable protein and the α/β hydrolase superfamily, containing the keratinase conserved domain; having obvious transmembrane structure and signal peptide cutting site, belonging to the secretory protein. In this study, we cloned the MGG-14095(38-281) gene of Magnaporthe oryzae, constructed the prokaryotic expression system pETM20-MGG_14095(38-281), used the stepwise chromatography to purify MGG_14095(38-281) protein, and the high-purity soluble target protein was obtained successfully. The results provided certain theoretical and experimental basis for the analysis of MGG_14095( 38-281) protein structure, the exploration and the control of pathogenetic mechanism of Magnaporthe oryzae.

参考文献

Ali Osman A.,and Munir T.,2017,Purification and characterization of cutinase from Bacillus sp.KY0701 isolated from plastic wastes,Prep.Biochem.Biotechnol.,47(9):925-933.

Bourett T.M.,and Howard R.J.,2011,In vitro development of penetration structures in the rice blast fungus Magnaporthe grisea,Can.J.Bot.,68(2):329-342.

Chen X.,Shi J.W.,Chen R.,Wen Y.A.,Shi Y.,Zhu Z.,Guo S.W.,and Li L.,2015,Molecular chaperones (TrxA,SUMO,Intein and GST) mediating expression,purification and antimicrobial activity assays of plectasin in Escherichia coli,Biotechnol.Appl.Biochem.,62(5):606-614.

Cheng Z.X.,Ren G.W.,Huang J.Y.,and Yang L.J.,2017,Synthesis of affinity ionic liquids for the purification of hexahistidine-tagged proteins,Zhongguo Shipin Xuebao (Journal of Chinese Institute of Food Science and Technology),17(12):122-128.(程志先,任广威,黄建颖,杨利军,2017,组氨酸标签蛋白纯化介质的合成及其分离纯化,中国食品学报,17(12):122-128.)

Chong F.C.,Tan W.S.,Biak D.R.A.,Ling T.C.,and Tey B.T.,2009,Purification of histidine-tagged nucleocapsid protein of Nipah virus using immobilized metal affinity chromatography,J.Chromatogr.B Analyt.Technol.Biomed.Life Sci.,877(14-15):1561-1567.

Deng Y.W.,Zhai K.R.,Xie Z.,Yang D.Y.,Zhu X.D.,Liu J.Z.,Wang X.,Qin P.,Yang Y.Z.,Zhang G.M.,Li Q.,Zhang J.F.,Wu S.Q.,Milazzo J.,Mao B.Z.,Wang E.T.,Xie H.A.,Tharreau D.,and He Z.H.,2017,Epigenetic regulation of antagonistic receptors confers rice blast resistance with yield balance,Science,355(6328):962-965.

Duan X.J.,Liu Y.,You X.,Jiang Z.Q.,Yang S.X.,and Yang S.Q.,2017,High-level expression and characterization of a novel cutinase from Malbranchea cinnamomea suitable for butyl butyrate production,Biotechnol.Biofuels.,10(1):223.

Fernandez J.,and Orth K.,2018,Rise of a cereal killer:the biology of Magnaporthe oryzae biotrophic growth,Trends Microbiol.,26(7):285-597.

Hamer J.E.,and Talbot N.J.,1998,Infection-related development in the rice blast fungus Magnaporthe grisea,Curr.Opin.Microbiol.,1(6):693-697.

Li X.C.,2018,Castor RcDELLA(GAI) protein expression,purification and crystal growth conditions filtering,Thesis for M.S.,Inner Mongolia University for Nationalities,Supervisor:Chen Y.S.,pp.22-28.(李晓晨,2018,蓖麻RcDELLA (GAI)蛋白的表达、纯化与晶体生长条件的筛选,硕士学位论文,内蒙古民族大学,导师:陈永胜,pp.22-28.)

Lu Z.Y.,Chen S.X.,Yao Y.Y.,Xing M.M.,and Xie Y.,2015,Research of protein separation and purification methods,Guangzhou Huagong (Guangzhou Chemical Industry),43(17):12-13,27.(卢忠英,陈仕学,姚元勇,邢明明,谢勇,2015,蛋白质分离纯化方法的研究进展,广州化工,43(17):12-13,27.)

Martin U.M.,Oses R.M.,Ryder L.S.,and Talbot N.J.,2016,Investigating the biology of plant infection by the rice blast fungus Magnaporthe oryzae,Fungal Genet.Biol.,90:61-68.

Qiu F.L.,and Wang D.W.,2004,Progress in nosogenesis of the rice blast fungus,Kenzhi yu Daozuo (Reclaiming and Rice Cultivation),(3):26-28.(邱福林,王大为,2004,稻瘟病菌致病机理的研究进展,垦殖与稻作,(3):26-28.)

Roychowdhury M.,Jia Y.L.,and Cartwright R.D.,2013,Structure,function,and co-evolution of rice blast resistance genes,Zuowu Xuebao (Acta Agronomica Sinica),38(3):381-393.(Roychowdhury M.,贾育林,Cartwright R.D.,2013,水稻抗稻瘟病基因的结构、功能和共同进化,作物学报,38(3):381-393.)

Ye X.,Sun Q.,and Liu Z.,2015,Progress on Magnaporthe oryzae infection process related to signaling pathways,Zhongguo Nongye Keji Daobao (Journal of Agricultural Science and Technology),17(1):87-94.(叶幸,孙群,刘柱,2015,稻瘟菌侵染过程相关信号通路研究进展,中国农业科技导报,17(1):87-94.)

Zhang N.,and Xu D.,2017,Advances of molecular biology techniques in rapid bacterial identification,Guoji Erkexue Zazhi(International Journal of Pediatrics),44(5):332-335.(张妮,徐丁,2017,分子生物学技术在病原菌快速检测中的应用进展,国际儿科学杂志,44(5):332-335.)

Zhang X.N.,Ran Q.Q.,and Zhnag X.J.,2013,Research and application progress on cutinase,Zhongguo Niangzao (China Brewing),32(11):11-17.(张效宁,冉琴琴,张学俊,2013,角质酶的研究与应用进展,中国酿造,32(11):11-17.)

基本信息:

DOI:10.13417/j.gab.040.001590

中图分类号:S435.111.41

引用信息:

[1]王双,李国瑞,陈永胜,等.稻瘟病菌MGG_14095基因克隆、表达及其表达产物的纯化[J].基因组学与应用生物学,2021,40(04):1590-1596.DOI:10.13417/j.gab.040.001590.

基金信息:

内蒙古民族大学科研项目(MDK2016008; MDK2018018; MDK2019020); 内蒙古自治区高等学校科学研究项目(NJZY20121)共同资助

发布时间:

2020-06-15

出版时间:

2020-06-15

网络发布时间:

2020-06-15

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