苏云金杆菌entomocidus亚种几丁质酶基因的克隆与生物信息学分析  

Cloning and Bioinformatics Analysis of the chitinase Gene from Bacillus thuringiensis Serovar entomocidus Strain HD109

作　　者：林毅[1] 彭锟[1] 关雄[2] 

机构地区：[1]华侨大学生物工程与技术系,福建泉州362021 [2]福建农林大学生物农药与化学生物学教育部重点实验室,福建福州350002

出　　处：《激光生物学报》2006年第6期598-601,共4页Acta Laser Biology Sinica

基　　金：国家自然科学基金(4060146);福建省高校新世纪优秀人才支持计划;福建省自然科学基金项目(B0510011);福建农林大学生物农药与化学生物学教育部重点实验室开放基金项目(KF0402)

摘　　要：从苏云金杆菌(Bacillus thuringiensis,B t)entom oc idus亚种HD109菌株中克隆了几丁质酶基因chiA74-HD109,其序列全长为2 031 bp,编码676个氨基酸,GenBank登录号为AY455290。其编码产物与蜡状芽孢杆菌几丁质酶CHiB(AB041932)的相似性达97.9%,与B t kenyae亚种LB IT-82菌株几丁质酶chiA74(AF424979)的相似性达98.4%,与B t pak istan i亚种几丁质酶(U89796)的相似性为83.0%,与B t kurstak i亚种几丁质酶kchi(AY189740)的相似性达97.8%,与B t israe lensis亚种几丁质酶(AF526379)的相似性达96.6%,与B t几丁质酶(AY074882)的相似性达98.4%,与B t sotto亚种几丁质酶(AY129671)的相似性为97.3%。功能结构域分析显示其编码区包含信号肽、催化区、Ⅲ型粘蛋白同源区及几丁质结合区4个部分。Based on the conserved 5 ＇ and 3＇ sequence of chitinase gene from B. thuringiensis, a product of 2 031 bp was amplified and cloned into Escherichia coli strain DHSot from B. thuringiensis serovar entomocidus strain HD109 genomic DNA. The product encoded an open reading frame （chiA74-HD109） encoding a deduced protein of 676 amino acids. Removal of the signal peptide sequence resulted in a predicted protein that was 70 457 Da in size and 5.45 of isoelectric point. The deduced 676 amino acids sequence showed high degree of identity with other chitinases such as ChiB （ AB041932 ） from Bacillus cereus （ 97.9 % ） , chiA74 （ AF424979 ） from B. thuringiensis serovar kenyae strain LBIT-82 （98.4 % ）, chitinase （U89796） from B. thuringiensis serovar pakistani （83.0 % ）, kchi （AY189740） from B. thuringiensis serovar kurstaki （ 97.8 % ） , chitinase （ AF526379 ） from B. thuringiensis serovar israelensis （ 96.6 % ） , chitinuse （AY074882） from B. thuringiensis （98.4 % ） , and chitinase （AY129671） from B. thuringiensis serovar sotto （97.3%）,Analysis of the sequence indicated that the chitinase contained a catalytie domain belonging to family 18 of glycosyl hydrolases in the N-terminus,a fibronectin type Ⅲ domain in the middle region and a chitin-binding domain in the - C-terminus,All three domains showed conserved sequences when compared to other bacterial chitinase sequences.

关 键 词：苏云金杆菌entomocidus亚种 几丁质酶 克隆 生物信息学分析 

分 类 号：Q932[生物学—微生物学]