农药学学报2012，Vol.14Issue(4)：398-404,7.DOI:10.3969/j.issn.1008-7303.2012.04.07
华北大黑鳃金龟Holotrichia oblita中肠羧酸酯酶基因HoCL1的克隆与序列分析
Molecular cloning and sequence analysis of HoCL1 carboxylesterase cDNAs from the midgut of Holotrichia oblita
摘要
Abstract
The peritrophic membrane protein polyclonal antiserum from Trichoplusia ni was used to screen cDNA expression library of the migut of Holotrichia oblita, and a cDNA clone encoding carboxylesterase named HoCLl was obtained. The sequence analysis indicated that the opening reading frame of HoCLl is 1599 bp in length encoding 532 amino acid residues of the proteinase with predicted molecular weight 59. 5 kDa and p/ 4. 5. HoCLl has conserved domains of carboxylesterases with a disulfide bond formation site,a serine activity center,only three cysteine residues and catalytic activity centers of Ser207, Asp333 and His422, and with no nitrogen linked glycosylation sites and oxygen glycosylation sites. Based on the amino acid sequence homology analysis and conserved domain analysis, HoCLl belongs to B esterase and is mostly similar to the carboxylesterase of Tribolium castaneum,but the similarity is only 35. 2%. In the sequence alignments and phylogenetic tree with other insect carboxylesterases, the sequence conservation of COOH-terminal amino acid in HoCLl is low,but the amino acid sequence near the N terminus is highly conserved. HoCLl clusters with the carboxylesterases of T. castaneum and Harmonia axyridis. Cloning and identification of HoCLl laid the foundation for the study of the expression and function in vivo.关键词
华北大黑鳃金龟/羧酸酯酶/基因克隆/序列分析/cDNA表达文库Key words
Holotrichia oblita/ carboxylesterase/ gene cloning/ sequence analysis/ cDNA expression library分类
生物科学引用本文复制引用
高勇,周洪旭,郑桂玲,李长友..华北大黑鳃金龟Holotrichia oblita中肠羧酸酯酶基因HoCL1的克隆与序列分析[J].农药学学报,2012,14(4):398-404,7.基金项目
山东省教育厅项目(J10LC07) （J10LC07）
青岛农业大学博士基金(6631024),"泰山学者"建设工程专项经费资助. （6631024）
