Bt菌株DQ89的sip基因的克隆、表达及杀虫活性分析  

Explore sip Gene from Bt Strain DQ89 and Its Insecticidal Activity against Colaphellus bowringi Baly

作　　者：张金波[1] 李海涛[1,2] 刘荣梅[1] 束长龙[2] 高继国[1] 

机构地区：[1]东北农业大学生命科学学院,哈尔滨150030 [2]中国农业科学院植物保护研究所/植物病虫害生物学国家重点实验室,北京100193

出　　处：《中国生物防治学报》2015年第4期598-602,共5页Chinese Journal of Biological Control

基　　金：转基因生物新品种培育科技重大专项(2014ZX0800913B-002);国家基础科学人才培养基金资助项目(J1210069);国家"863"计划(2011AA10A203);国家自然科学基金(31401812);黑龙江省教育厅科学技术研究项目(12521021);植物病虫害生物学国家重点实验室开放基金(SKLOF201405)

摘　　要：苏云金芽胞杆菌Bacillus thuringicnsis(Bt)是一种广泛存在于自然界中的生防微生物,其杀虫活性主要来源于杀虫基因编码的内毒素(insecticidal crystal proteins,ICPs)、营养期杀虫蛋白(vegetative insecticidal protein,VIP)和分泌期杀虫蛋白(secreted insecticidal protein,Sip),相比于以上2种杀虫蛋白,Sip的研究相对较少。本研究从实验室中的Bt野生菌株中克隆sip基因,其中编号为DQ89的Bt菌株成功克隆得到1188 bp大小的sip基因,编码395个氨基酸,与已知的Sip1A蛋白的相似性为87%,序列提交到Gen Bank并获得登录号KP114614。将该基因在大肠杆菌中表达的蛋白进行生物活性测定,结果显示其对大猿叶甲Colaphellus bowringi Baly具有一定的杀虫活性,其LC50为1.542μg/m L。Bacillus thuringiensis insecticidal crystal secreted insecticidal protein （SIP） were widely used proteins （ICPs）, vegetative insecticidal protein （VIP） and as a kind of safe bio-insecticides. ICP and VIP have been reported widely, while few studies have concerned on SIP. In this study, PCR method was used to clone and identify the sip gene from Bt strains isolated from different environments. A sip gene was obtained from the strain DQ89, which is of 1188 bp and encodes 395 amino acids, showing a 87% similarity of amino acid residues with the registered SiplA protein. The nucleotide sequencing was submitted to GenBank with an Accession No. of KP 114614. The protein expressed in E. coli showed an insecticidal activity against Colaphellus bowringi Baly, with LCs0 of 1.542 μg/mL.

关 键 词：苏云金芽胞杆菌 sip基因 生物活性 大猿叶甲 

分 类 号：S476.11[农业科学—农业昆虫与害虫防治]