中国人兽共患病学报2012，Vol.28Issue(6)：570-573,4.DOI:10.3969/cjz.j.issn.1002-2694.2012.06.011
家蝇几丁质酶基因的序列分析、克隆和诱导表达
Sequence analysis, cloning and induced expression of chitinaseⅠ gene in housefly(Musca domestica)
摘要
Abstract
The aim of this study is to analyze and predict the structure and characteristics of genes and encoding proteins of MDC Ⅰ (Musca domestica chitinase Ⅰ ), with the methods of cloning and expressing that gene. Sequence analysis revealed that the open reading frame of the cDNA encoded a 251-amino acid protein, which contained an NH2-terminal signal sequence (1-22). The sequence identified with other insect chitinase was between 60% and 70%. The protein, with a predicted molecular weight of 28. 62 kDa and pi of 5. 78, had one active site of family 18 chitinase. The gene coding for MDC I was amplified by polymerase chain reaction (PCR), and then was inserted into vector pET 28a ( + ) and induced with IPTG. The recombinant protein in the expression vector was analyzed by SDA-PAGE. Results indicated that the recombinant plasmid with the correct target gene was constructed, and the recombinant protein was expressed in E. coli BL21 (DE3). The target gene was cloned into the host bacterium and expressed correctly, and these results would establish the basis for further researches in biology and immunology of chitinase in housefly.关键词
家蝇/几丁质酶/序列分析/基因克隆/表达Key words
Musca domestica/ chitinase/ sequence analysis/ gene cloning/ expression分类
医药卫生引用本文复制引用
国果,吴建伟,吴沁怡,付萍,张勇..家蝇几丁质酶基因的序列分析、克隆和诱导表达[J].中国人兽共患病学报,2012,28(6):570-573,4.基金项目
国家自然科学基金(81160204,30960343) （81160204,30960343）
贵州省科技厅基金(黔科合[2010]3160) （黔科合[2010]3160）
高校博士点学科专项科研基金(20105215120001) （20105215120001）
