拟环纹豹蛛热休克蛋白20基因的克隆与分析  

Cloning and analysis of Pardosa pseudoannulata heat shock protein 20 gene

作　　者：李长春 王永[1,2] 姚国新 戴余军[1,2] 李国元 LI Chang-chun;WANG Yong;YAO Guo-xin;DAI Yu-jun;LI Guo-yuan(Hubei Key Laboratory of Quality Control of Characteristic Fruits and Vegetables,Hubei Engineering University,Xiaogan,Hubei 432000,China;College of Life Science and Technology,Hubei Engineering University,Xiaogan,Hubei 432000,China)

机构地区：[1]湖北工程学院特色果蔬质量安全控制湖北省重点实验室,湖北孝感432000 [2]湖北工程学院生命科学技术学院,湖北孝感432000

出　　处：《南方农业学报》2018年第8期1525-1530,共6页Journal of Southern Agriculture

基　　金：湖北省自然科学基金面上项目(2017CFB609);区域开发与环境响应湖北省重点实验室开放基金项目(2016C002);特色果蔬质量安全控制湖北省重点实验室开放基金项目(2017K02)

摘　　要：【目的】克隆拟环纹豹蛛热休克蛋白20基因(HSP20),并分析其在镉胁迫下的表达情况,为揭示蜘蛛抵御重金属胁迫机制提供参考。【方法】以拟环纹豹蛛雌蛛为试验材料,在转录组测序的基础上,通过RT-PCR克隆拟环纹豹蛛HSP20基因(PpHSP20),用生物信息学方法对其序列特征进行分析,并采用荧光定量PCR检测镉胁迫下PpHSP20基因的相对表达量。【结果】克隆获得的PpHSP20基因编码区长525 bp,编码174个氨基酸,其编码蛋白分子量20.08kD,等电点5.97,具有小分子热休克蛋白家族典型的α-晶状体蛋白结构域,与温室拟肥腹蛛(Parasteatoda tepidariorum)α-晶体蛋白A有较高的相似性(58%)。荧光定量PCR分析结果显示,镉胁迫下PpHSP20基因的表达量显著增加(P<0.05)。【结论】拟环纹豹蛛HSP20基因在抵御镉胁迫中发挥调控作用,可作为研究蜘蛛抵御镉胁迫的重要候选基因和潜在的生物标记物。【Objective】In order to provide some references for revealing the mechanisms by which spiders resisted to heavy metals,the small heat shock protein 20 gene(HSP20)of Pardosa pseudoannulata was cloned and its expression under cadmium was analyzed.【Method】Female P.pseudoannulata was used as materials,HSP20 gene of the P.pseudoannulata(PpHSP20)was cloned using RT-PCR technique based on the data obtained from transcriptome sequencing.PpHSP20 sequence characters were analyzed by bioinformatics method.The relative expression levels of PpHSP20 under cadmium stress were investigated by fluorescence quantitative PCR.【Result】The encoding region of the HSP20 gene was 525 bp in length,encoding 174 amino acids.Its molecular weight of encoded protein was 20.08 kD.It had 5.97 theoretical isoelectric point and typicalα-crystallin domain of small heat shock proteinfamily.The deduced amino acid sequence of PpHSP20 shared high homology withα-crystallin of Parasteatoda tepidariorum(58%).Fluorescence quantitative PCR indicated that the expression of gene PpHSP20 under cadium stress increased significantly(P<0.05).【Conclusion】Gene PpHSP20 plays a regulation role in resistance to cadium stress,and can be an important candidate gene in research of spiders resisting cadium stress and a potential marker.

关 键 词：拟环纹豹蛛 HSP20基因 基因克隆 荧光定量PCR 

分 类 号：S186[农业科学—农业基础科学] Q956[生物学—动物学]