柑橘几丁质酶Cs1g26330.1基因克隆及原核表达  

Cloning and Prokaryotic Expression of Citrus Chitinase Cs1g26330.1 Gene

作　　者：胡亚平 庄秋帆[1] 吉前华 郭雁君[1,2] 蒋惠[1,2] 郭丽英 Hu Yaping;Zhuang Qiufan;Ji Qianhua;Guo Yanjun;Jiang Hui;Guo Liying(Institute of Fruit Tree Research,Zhaoqing University,Zhaoqing,526061;School of Life Sciences,Zhaoqing University,Zhaoqing,526061)

机构地区：[1]肇庆学院果树研究所,肇庆526061 [2]肇庆学院生命科学学院,肇庆526061

出　　处：《分子植物育种》2024年第19期6295-6300,共6页Molecular Plant Breeding

基　　金：现代农业产业技术体系建设专项(CARS-26);广东省自然科学基金项目(2016A030313014);广东省大学生创新创业训练计划项目(S202010580041)共同资助。

摘　　要：几丁质酶具有糖苷水解酶和溶菌酶活性,能降解几丁质,是植物重要的抗真菌病基因。为了研究柑橘几丁质酶的催化功能,了解其编码蛋白质的性质,为后续抗病育种研究打下基础。本研究从砂糖橘cDNA文库中克隆了几丁质酶Cs1g26330.1基因的全长编码区序列,将基因连接到pEASY-Blunt E1载体中,测序验证后,提取重组质粒并转化E.coli BL21(DE3)感受态细胞,以IPTG诱导该基因成功表达,经SDS-PAGE电泳检测到重组蛋白质。本研究获得的在大肠埃希菌中表达几丁质酶可以作为蛋白抗原去开发特异性抗体,用来深入研究柑橘几丁质酶的功能,也有望作为植物抗病的高效无毒农药使用,或制作降解酶用于几丁质的回收利用。Chitinase has glycoside hydrolase and lysozyme activities,can degrades chitin,and is an important fungal disease resistance gene in plants.In order to study the catalytic function of chitinases in citru and understand the properties of the proteins,and lay a foundation for the subsequent research on disease resistance breeding.In this study,the full-length coding region sequence of chitinase gene Cs1g26330.1 was amplified from a cDNA library of Citrus Shatangju,then cloned into the pEASY-Blunt E1 vector,after sequenced and verified,the recombinant plasmid was transformed E.coli BL21(DE3)compendent cells,the gene was expression successfully induced by IPTG,and the recombinant protein was detected by SDS-PAGE electrophoresis.The chitinase expression in E.coli obtained in this study can be used as a protein antigen to develop specific antibodies,which can be used for further study the function of Citrus chitinase,and is also expected to be used as an efficient and non-toxic pesticide for plant disease resistance,or to produce degradative enzymes for utilization of chitin.

关 键 词：柑橘 几丁质 几丁质酶 原核表达 重组蛋白 

分 类 号：S666[农业科学—果树学]