生物技术通报2016，Vol.32Issue(3)：115-121,7.DOI:10.13560/j.cnki.biotech.bull.1985.2016.03.019
家蝇伴侣蛋白CCTδ基因克隆、序列分析及表达模式的研究
Cloning,Bioinformatic Analysis and Expression Patterns of Chaperonin Gene CCTδ fromMusca domestica
摘要
Abstract
This work aims to clone,have bioinformatic analysis and explore the expression patterns of chaperonin gene CCTδfromMusca domestica. We screened and isolated the gene CCTδ from the built cDNA plasmid library ofM. domesticalarva by EST sequencing,and having the plasmid as template,the amplification was conducted by PCR. Further,with bioinformatics we analyzed the structures and functions of gene CCTδ and the encoded protein,and constructed phylogenetic tree. We collected the bodies at different developmental stages ofM. domestica life history(eggs,varied-instar larvae,pupae,female and male adults),and chose 6 tissues of 3-instar larvae(body wall,trachea,salivary glands,fat body,markov tube,and midgut),thus the expression patterns of gene CCTδ at different developmental stages and tissues ofM. domestica were detected by real-time quantitative PCR. An about 1 600 bp specific fragment of housefly’s gene CCTδ was acquired by PCR amplification. The open reading frame of gene CCTδ was 1 602 bp that encoded a putative protein with 533 amino acids. The predicted molecular weight of the protein was 57.16 kD and pI was 7.50. The secondary structures were mainly composed ofα-helix and random coil. Comparative analysis of the phylogenetic tree showed that it was conservative,and the genetic distance was closer withAnopheles darlingi. The spatial and temporal expression patterns of geneCCTδ revealed that gene CCTδ expressed in each developmental stage ofM. domestica,while the highest at pupa stage;and it expressed the highest in trachea among various tissues of 3-instar larvae. Conclusively,in this study,the gene CCTδ of M. domesticawas successfully cloned,and the spatial and temporal expression patterns of gene CCTδ were preliminarily explored.关键词
家蝇/CCTδ基因/生物信息学分析/表达模式Key words
Musca domestica/CCTδ gene/bioinformatic analysis/expression pattern引用本文复制引用
陶如玉,赵学军,杨尉锦,吴建伟,国果..家蝇伴侣蛋白CCTδ基因克隆、序列分析及表达模式的研究[J].生物技术通报,2016,32(3):115-121,7.基金项目
国家自然科学基金项目（81160204，81360254），贵州省卫生厅基金项目（gzwjkj2014-2-100），国家科技部支撑计划课题子课题（2011BAC06B12），贵州省科学技术基金项目 （）
