摘要
柑橘黄龙病是我国柑橘产业可持续发展面临的重大威胁,建立高效、准确、灵敏的检测方法是黄龙病防控的关键.通过生物信息学分析柑橘黄龙病菌亚洲种(CLas)不同菌株基因组,预测出83个经典分泌蛋白和87个非经典分泌蛋白.选定其中14个分泌蛋白基因,经大肠杆菌密码子优化后构建pET-28a-CLasSec和pCold-I-CLasSec系列原核表达载体.经IPTG诱导条件优化、镍磁珠法纯化后,获得了CLasSec2、CLasSec4、CLasSec5、CLasSec6、CLasSec8和CLasSec12等6个柑橘黄龙病菌亚洲种分泌蛋白.
An efficient, accurate and sensitive detection tool for citrus Huanglongbing(HLB) is considered a critical step for the sustainability of citrus industry. To establish a detection method, 83 classical and 87 non-classical secretory proteins were predicted from ‘Candidatus Liberibacter asiaticus’(CLas) genome through bioinformatics analysis. Then 14 candidate genes were firstly optimized by Escherichia coli codon and then cloned into pET-28 a or pCold-I vector for prokaryotic expression. After optimization by IPTG induction and protein purification using Ni-Charged MagBeads, 6 secretory proteins of CLasSec2, CLasSec4, CLasSec5, CLasSec6, CLasSec8 and CLasSec12 were successfully obtained.
作者
王春庆
钟八莲
卢占军
黄爱军
陈慈相
苏华楠
WANG Chunqing;ZHONG Balian;LU Zhanjun;HUANG Aijun;CHEN Cixiang;SU Huanan(National Navel Orange Engineering Research Center/School of Life Science,Gannan Normal University;Fruit Bureau of Ganzhou,Ganzhou,Jiangxi 341000,China)
出处
《福建农林大学学报（自然科学版）》
CSCD
北大核心
2020年第3期313-319,共7页
Journal of Fujian Agriculture and Forestry University:Natural Science Edition
基金
江西省科技计划项目(20143ACF60012)
国家重点研发计划(2018YFD0201500)
江西省现代农业产业技术体系建设专项资金项目(JXARS-07)
赣州市科技计划项目(赣市财教字[2017]8号)
江西省研究生创新专项资金项目(YC2018-S394).
关键词
柑橘黄龙病
分泌蛋白
效应子
原核表达
citrus Huanglongbing
secretory protein
effector
prokaryotic expression
