甘蔗谷胱甘肽硫转移酶基因SoGST-1a的克隆与表达分析

doi:10.11924/j.issn.1000-6850.casb15040019

中国农学通报 ›› 2015, Vol. 31 ›› Issue (26): 71-77.doi: 10.11924/j.issn.1000-6850.casb15040019

所属专题：生物技术

甘蔗谷胱甘肽硫转移酶基因SoGST-1a的克隆与表达分析

徐磊,姚艳丽,胡小文,邢淑莲,刘洋

(中国热带农业科学院湛江实验站,广东湛江 524013）

收稿日期:2015-04-02 修回日期:2015-05-20 接受日期:2015-05-25 出版日期:2015-09-23 发布日期:2015-09-23
通讯作者: 刘洋
基金资助:
中央级公益性科研院所基本科研业务费专项“甘蔗抗氧化系统关键酶基因的克隆与表达分析”(ITBB120504)；中国热带农业科学院湛江实验站科研启动专项资金项目“高粱全基因组WRKY转录因子的鉴定、表达分析及克隆”(zjky201504)。

Cloning and Expression Analysis of Glutathione S-transferase Gene SoGST-1a in Sugarcane

Xu Lei, Yao Yanli, Hu Xiaowen, Xing Shulian, Liu Yang

(Zhanjiang Experiment Station, Chinese Academy of Tropical Agricultural Sciences, Zhanjiang Guangdong 524013)

Received:2015-04-02 Revised:2015-05-20 Accepted:2015-05-25 Online:2015-09-23 Published:2015-09-23

摘要/Abstract

摘要： 克隆甘蔗谷胱甘肽硫转移酶(Glutathione S-transferase，GST)基因，并分析其序列特征和表达谱，为甘蔗抗旱育种提供基因来源。本研究采用电子克隆和RT-PCR技术从甘蔗品种‘新台糖22号’中克隆到一个甘蔗GST基因，命名为SoGST-1a。通过生物信息学软件分析其序列特征，采用实时荧光定量PCR检测其表达谱。序列分析表明该基因cDNA全长702 bp，编码224个氨基酸，预测的蛋白分子式为C₁₁₁₇H₁₇₅₀N₃₀₀O₃₂₈S₆，蛋白分子量为24.82 kDa，等电点为5.96。蛋白疏水性分析推测其为亲水性蛋白。保守结构域预测表明SoGST-1a蛋白具有2个GST结构域。系统进化树分析表明该基因与玉米Zeta类GST蛋白亲缘关系最近。荧光定量PCR分析显示，干旱胁迫下SoGST-1a基因下调表达。研究结果表明，SoGST-1a基因可能在甘蔗干旱胁迫中并不发挥一定作用。这些结果为深入了解SoGST-1a基因的生物学功能奠定了基础。

关键词: 农业信息网站, 农业信息网站, 信息服务质量评价, 优化, R型聚类, 多元线性回归

Abstract: The glutathione S-transferase (GST) gene from sugarcane was cloned. Its sequence characteristics and expression patterns were analyzed. This provided the source of gene in sugarcane breeding for drought resistance. In this study, a GST gene from sugarcane variety ROC22 was cloned by in silico cloning and RT-PCR, named SoGST-1a. Its sequence characteristics were analyzed by using bioinformatics software, and its expression patterns were detected by using real-time PCR. Sequence analysis showed that the full length cDNA of SoGST-1a was 702 bp, which encoded 224 amino acids. The predicted protein molecular formula was C₁₁₁₇H₁₇₅₀N₃₀₀O₃₂₈S₆, whose molecular mass was 24.82 kDa and isoelectric point was 5.96. Hydrophobic analysis indicated that it was a hydrophilic protein. Conserved domain prediction showed that SoGST-1a protein had?two GST domains. Phylogenetic analysis showed that SoGST-1a protein was close to Zeta class GST protein of maize. Real-time PCR results showed that the SoGST-1a gene was down-regulated expression under drought treatment. The research results showed that SoGST-1a gene might not play a role in drought stress in sugarcane. These results provided a basis for further understanding biological function of SoGST-1a.

徐磊,姚艳丽,胡小文,邢淑莲,刘洋. 甘蔗谷胱甘肽硫转移酶基因SoGST-1a的克隆与表达分析[J]. 中国农学通报, 2015, 31(26): 71-77.

Xu Lei,Yao Yanli,Hu Xiaowen,Xing Shulian and Liu Yang. Cloning and Expression Analysis of Glutathione S-transferase Gene SoGST-1a in Sugarcane[J]. Chinese Agricultural Science Bulletin, 2015, 31(26): 71-77.

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甘蔗谷胱甘肽硫转移酶基因SoGST-1a的克隆与表达分析

Cloning and Expression Analysis of Glutathione S-transferase Gene SoGST-1a in Sugarcane

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