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
胡亚平 1庄秋帆 2吉前华 1郭雁君 1蒋惠 1郭丽英1
作者信息
- 1. 肇庆学院果树研究所,肇庆,526061;肇庆学院生命科学学院,肇庆,526061
- 2. 肇庆学院果树研究所,肇庆,526061
- 折叠
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.
Abstract
Chitinase has glycoside hydrolase and lysozyme activities,can degrades chitin,and is an important fun-gal 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 li-brary of Citrus Shatangju,then cloned into the pEASY-Blunt E1 vector,after sequenced and verified,the recombi-nant 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Key words
Citrus/Chitin/Chitinase/Prokaryotic expression/Recombinant protein引用本文复制引用
基金项目
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)
出版年
2024
NETL
NSTL
万方数据