畜牧与兽医2017，Vol.49Issue(11)：118-123,6.
少孢节丛孢菌新疆分离株几丁质酶AO-14基因的克隆表达
Cloning and expression of chitinase AO-14 gene from Arthrobotrys oligospora XJ-A1
摘要
Abstract
The target gene fragment of chitinase A0-14 of Arthrobotrys oligospora XJ-A1 was obtained by RT-PCR.The gene was cloned and its molecular characterizations were analyzed.The recombinant plasmid pET32a-AO14 was transformed into Escherichia coli BL21 and induced by IPTG.The recombinant protein was identified by SDS-PAGE and Western blot.The results showed that the full length of AO-14 gene was 1200bp,encoding 399 amino acids.Chitinase AO-14 belongs to the family of glycoside hydrolase 18,which is characterized by a typical chitinase catalytic region with the conserved sequence SXGGDGXDXDWE,amino acid sequence of 131 to 139 as the active site area and no signal peptide sequence.SDS-PAGE analysis showed that the protein product had a molecular weight of about 60 ku.Western blot analysis indicated that the AO-14 recombinant protein could specifically react with polyclonal antibody against protein crude extracts.This study provides a basis for investigation of chitinase AO-14 function.关键词
少孢节丛孢菌/几丁质酶/AO-14基因/克隆/序列分析/原核表达Key words
Arthrobotrys oligospora/chitinase/AO-14 gene/cloning/sequence analysis/prokaryotic expression分类
农业科技引用本文复制引用
钟文强,孟庆玲,乔军,陈英,贡莎莎,才学鹏..少孢节丛孢菌新疆分离株几丁质酶AO-14基因的克隆表达[J].畜牧与兽医,2017,49(11):118-123,6.基金项目
国家自然科学基金(31460654,31260601) （31460654,31260601）
新疆自治区研究生科研创新项目(XJGRI2015038) （XJGRI2015038）
