Cloning, Bioinformatic Analysis and Expression Patterns of Chaperonin Gene CCTδ from Musca domestica

doi:10.13560/j.cnki.biotech.bull.1985.2016.03.019

Abstract

Abstract: This work aims to clone, have bioinformatic analysis and explore the expression patterns of chaperonin gene CCTδ from Musca domestica. We screened and isolated the gene CCTδ from the built cDNA plasmid library of M. domestica larva 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 of M. 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 of M. 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 of α-helix and random coil. Comparative analysis of the phylogenetic tree showed that it was conservative, and the genetic distance was closer with Anopheles darlingi. The spatial and temporal expression patterns of gene CCTδ revealed that gene CCTδ expressed in each developmental stage of M. 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. domestica was successfully cloned, and the spatial and temporal expression patterns of gene CCTδ were preliminarily explored.

Key words: Musca domestica, CCTδ gene, bioinfor matic analysis, expression pattern

TAO Ru-yu, ZHAO Xue-jun, YANG Yu-jin, WU Jian-wei, GUO Guo. Cloning, Bioinformatic Analysis and Expression Patterns of Chaperonin Gene CCTδ from Musca domestica[J]. Biotechnology Bulletin, 2016, 32(3): 115-121.

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