苏云金芽孢杆菌LLP29菌株中ykkB基因的克隆、表达及活性分析  

Cloning,expression and activity analysis of ykkB in Bacillus thuringiensis LLP29

作　　者：杨兆辉 汪子洋 杨文涛 蔡瑜婷 谭伟龙 黄恩炯 张灵玲[1] YANG Zhaohui;WANG Ziyang;YANG Wentao;CAI Yuting;TANG Weilong;HUANG Enjiong;ZHANG Lingling(College of Life Sciences,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China;Center for Disease Control and Prevention of Eastern Command,Nanjing,Jiangsu 210002,China;Fuzhou International Travel Healthcare Center,Fuzhou,Fujian 350001,China)

机构地区：[1]福建农林大学生命科学学院,福建福州350002 [2]东部战区疾病预防控制中心,江苏南京210002 [3]福州国际旅行卫生保健中心,福建福州350001

出　　处：《福建农林大学学报（自然科学版）》2022年第1期33-38,共6页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：福建省自然科学基金(2020J01550、2020I0031);福建省卫生联合攻关计划(2019-WJ-29);生物农药与化学生物学教育部重点实验室开放课题(Keylab2018-04);闽台合作有害生物生态防控国家重点实验室开放课题(SKL2019007);害虫绿色防控福建省高等学校重点实验室开放研究基金(PTJH1700701);福建农林大学科技创新专项基金(KFA20124A).

摘　　要：乙酰化是蛋白质翻译后修饰的一种重要方式,可参与多种细胞进程,但迄今未见有关乙酰化修饰在Bt中发挥作用的报道.本课题组前期通过对高效杀虫Bt菌株LLP29的基因组进行测序分析,发现该菌株也存在编码乙酰转移酶的基因(如ykkB).为探讨乙酰化修饰在Bt菌株中的作用,以ykkB为靶标,利用无缝克隆技术成功克隆与测序了ykkB基因.通过生物信息学分析,发现该基因编码的蛋白可能是一类N-乙酰转移酶,赖氨酸和亮氨酸含量最高,半胱氨酸含量最低,是一种亲水性蛋白质,且无跨膜区;同时,成功表达、纯化了YkkB蛋白,通过体外自乙酰化分析,证实该蛋白可发挥乙酰基转移的作用.As an important method in protein post-translational modification(PTM),acetylation participates in various cellular processes.However,the role of acetylation modification in Bt remains unknown.In our previous work,ykkB,a type of acetyltransferase gene,was detected in the Bt strain LLP29 by genome sequencing.To further explore the role of acetylation modification in Bt strain,ykkB was cloned and sequenced again by seamless cloning technology.Bioinformatics analysis showed that YkkB might be a kind of N-acetyltransferase,with the highest lysine and leucine content and the lowest cysteine content.It is a hydrophilic protein without transmembrane region.After being expressed and purified,YkkB was confirmed to regulate acetyl transfer by autoacetylation analysis in vitro.

关 键 词：BT ykkB基因 克隆 表观遗传 乙酰化 

分 类 号：S476[农业科学—农业昆虫与害虫防治] Q785[农业科学—植物保护]