家蝇几丁质酶基因的鉴定与表达分析  

Identification and expression analysis of chitinase gene in Musca domestica

作　　者：赵文静 焦正龙 苏佩佩 尚小丽[2] 修江帆 国果 ZHAO Wen-jing;JIAO Zheng-long;SU Pei-pei;SHANG Xiao-li;XIU Jiang-fan;GUO Guo(School of Basic Medical Sciences,Guizhou Medical University,Guiyang 550025,China;Key Laboratory of Environmental Pollution Monitoring and Disease Control;Translational Medicine Research Center of Guizhou Medical University)

机构地区：[1]贵州医科大学基础医学院,贵州贵阳550025 [2]贵州医科大学环境与健康重点实验室 [3]贵州医科大学转化医学研究中心

出　　处：《中国病原生物学杂志》2023年第4期432-438,共7页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.81560337,81760647)。

摘　　要：目的从基因家族的角度对家蝇几丁质酶进行鉴定与分析,初步探讨其潜在的生物学功能。方法采用生物信息学方法分析家蝇几丁质酶基因的结构、组成及蛋白的理化特性,并构建分子系统树;通过qPCR检测8个基因的时空表达情况和4个基因在3种浓度的20E处理下的表达变化。结果共鉴定20个家蝇几丁质酶基因,N-J树划分为8组;不同基因的外显子数量及结构域组成存在差异性;催化结构域具备昆虫几丁质酶高保守序列;相对分子质量为(26.663~270.602)×10^(3),pI为5.02~10.28。MdCht2、MdCht5、MdCht7和MdCht10在蛹期显著性高表达;MdCht4、MdCht8、MdCht9和MdIDGF7主要在幼虫期高表达;Ⅱ龄幼虫的8种几丁质酶基因呈组织特异性表达,其中在马氏管和肠组织中MdCht4显著性高表达。MdCht4、MdCht5、MdCht7和MdCht10对蜕皮激素20E的刺激呈正向应答,注射后8h和12h时的应激表达量与20E的浓度相关。结论家蝇丁质酶基因呈组织特异性表达,部分几丁质酶基因对蜕皮激素20E的刺激呈正向应答,本文可为家蝇几丁质酶的功能研究及家蝇的防治提供参考。Objective This study aimed to investigate the potential biological function of insect chitinase by analyzing the chitinase gene family in Musca domestica.Methods Bioinformatics methods were used to describe the structures,composition and physicochemical characteristics of housefly chitinases.qPCR was used to detect the expression patterns of eight chitinases.Results Atotal of twenty chitinase genes were identified and divided into eight groups by N-J phylogenetic tree.Each chitinase displayed a distinct schematic diagram of exon and intron organization and domain organization.The catalytic domains of insect chitinase have highly conserved sequences,the molecular weight ranged from(26.663-270.602)×10^(3),while the pI ranged from 5.02-10.28.During the pupal stage,MdCht2(F_(6,12)=72.86),MdCht5(F_(6,12)=662.5),MdCht7(F_(6,12)=72.86)and MdCht10(F_(6,12)=261.6)were significantly expressed,while MdCht4(F_(6,12)=114.3),MdCht8(F_(6,12)=33.73),MdCht9(F_(6,12)=445.2)and MdIDGF7(F_(6,12)=71.38)were significantly expressed mainly in the larval stage(P0.01).Eight chitinase genes showed tissue-specific expression in the 2nd-instar larvae stage.In Malpighian tube and intestinal tissue,MdCht4 was significantly expressed(F_(4,8)=180.6 and F_(4,8)=112.8,P0.01).Besides,MdCht4,MdCht5,MdCht7 and MdCht10 responded positively to the stimulation of ecdysone 20E,and their stress expression levels correlated with 20E concentration at 8 h and 12 h after injection.Conclusion Our results demonstrate that chitinase genes of M.domestica exhibited tissue-specific expression patterns,and some chitinase genes responded positively to the stimulation of ecdysone 20E.This work may provide a reference for the functional study of housefly chitinase and the control of housefly.

关 键 词：几丁质酶 基因家族 表达模式 20-羟基蜕皮激素诱导(20E) 家蝇 

分 类 号：R384.2[医药卫生—医学寄生虫学]