[1]李长春,宁青,戴余军,等.拟环纹豹蛛谷胱甘肽S-转移酶基因的克隆及表达分析[J].江苏农业学报,2019,(05):1068-1074.[doi:doi:10.3969/j.issn.1000-4440.2019.05.010]
　LI Chang-chun,NING Qing,DAI Yu-jun,et al.Cloning and expression analysis of glutathione S-transferase gene in Pardosa pseudoannulata[J].,2019,(05):1068-1074.[doi:doi:10.3969/j.issn.1000-4440.2019.05.010]
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拟环纹豹蛛谷胱甘肽S-转移酶基因的克隆及表达分析()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2019年05期
页码:
1068-1074
栏目:
植物保护
出版日期:
2019-10-31

文章信息/Info

Title:
Cloning and expression analysis of glutathione S-transferase gene in Pardosa pseudoannulata
作者:
李长春1; 宁青1; 2; 戴余军1; 王立华1; 李国元1; 彭宇2
(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．湖北大学生命科学学院省部共建生物催化与酶工程国家重点实验室，湖北武汉430062)
Author(s):
LI Chang-chun1; NING Qing1; 2; DAI Yu-jun1; WANG Li-hua1; LI Guo-yuan1; PENG Yu2
(1.Hubei Key Laboratory of Quality Control of Characteristic Fruits and Vegetables, Hubei Engineering University, Xiaogan 432000, China;2.The State Key Laboratory of Biocatalysis and Enzyme Engineering of China, College of Life Sciences, Hubei University, Wuhan 430062, China)
关键词:
拟环纹豹蛛; 谷胱甘肽S-转移酶; 基因克隆; 表达分析
Keywords:
Pardosa pseudoannulata; glutathione S-transferase; gene cloning; expression analysis
分类号:
Q785
DOI:
doi:10.3969/j.issn.1000-4440.2019.05.010
文献标志码:
A
摘要:
为研究拟环纹豹蛛谷胱甘肽S-转移酶(GST)基因碱基序列及其对镉胁迫的响应，本研究在转录组测序的基础上，通过RT-PCR克隆了拟环纹豹蛛的GST基因(PpGST)，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检测了镉胁迫下PpGST基因的相对表达量。结果表明，克隆获得的PpGST(GenBank登录号为KY454857)基因编码区长654 bp，可编码1个217个氨基酸的蛋白质，该蛋白质理论分子量为24 900，等电点为5.98，具有GST蛋白家族保守的N端结构域和C端结构域，与温室拟肥腹蛛(Parasteatoda tepidariorum)GST蛋白Delta型有较高的相似性(65%)。荧光定量PCR分析结果显示，镉胁迫下PpGST基因的表达量显著增加(P<0.05)，暗示其在抵御镉胁迫中可能发挥了重要作用。
Abstract:
To analyze glutathione S-transferase (GST) gene sequence of the wolf spider Pardosa pseudoannulata and its response to cadmium(Cd) stress#, GST gene of the P. pseudoannulata (PpGST) was cloned using RT-PCR technique, based on the transcriptomic data. The sequence characteristics of PpGST gene were analyzed by bioinformatics. The relative expression levels of PpGST gene under cadmium stress were investigated by real-time fluorescent quantitative RT-PCR. The coding region of the PpGST gene (GenBank accession No. KY454857) was 654 bp in length, and the gene could encode a protein of 217 amino acid residues. The theoretical molecular weight of the protein was 24 900, and the isoelectric point was 5.98. The predicted protein contained the conservative N-terminal domain and C-terminal domain of the GST protein family. The deduced amino acid sequence of PpGST shared high homology with GST Delta of Parasteatodatepidariorum(65%). Fluorescence quantitative PCR analysis results showed that the expression level of PpGST increased significantly under Cd stress, suggesting that it might play an important role in response to Cd stress.

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备注/Memo

备注/Memo:
收稿日期：2019-01-28 基金项目：国家自然科学基金项目(31672317)；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(湖北大学)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(湖北工程学院)开放基金项目(2017K02) 作者简介：李长春(1976-)，男，湖北广水人，博士，副教授，主要从事动物毒理学研究。(E-mail) lcc386@163.com 通讯作者：彭宇，(E-mail) pengyu@hubu.edu.cn
更新日期/Last Update: 2019-11-11