Cloning and expression of a tomato glutathione S- transferase (GST) in Escherichia coli

Dongli Pei; Jinping Ding; Zilei Duan; Meng Li; Yiyang Feng; Chengwei Li

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Cloning and expression of a tomato glutathione S- transferase (GST) in Escherichia coli

机译：番茄谷胱甘肽S-转移酶（GST）的克隆及在大肠杆菌中的表达

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Glutathione S- transferases (GSTs) fulfill a diverse range of functions in an organism. In a previous study, a putative glutathione S-transferase gene (ShGSTU1) from a wild-type tomato,?Solanum?habrochaites?G1.1560, was identified to be a key gene in pathogen resistant response against powdery mildew in tomato. In this study,ShGSTU1?was cloned into plasmid pET-28a, efficiently expressed in?Escherichia coliupon isopropyl-β-D-1-thiogalactopyronoside (IPTG) induction, purified with Ni2+?affinity chromatography and biochemically characterized. The results show that the optimal conditions for the expression of recombinant ShGSTU1 in?E. coli?were growth under 37°C, and 4-h IPTG induction with 1 mM concentration. About 18.93 mg ShGSTU1 was recovered from 1 g wet bacteria. The recombinant ShGSTU1 exhibited enzymatic activity with specific activity 0.625 U/mg. These results might provide a significant foundation for the later research on the mechanism of ShGSTU1 in tomato resistance to powdery mildew.

机译：谷胱甘肽S-转移酶（GST）在生物体中具有多种功能。在先前的研究中，来自野生型番茄的一个推定的谷胱甘肽S-转移酶基因（ShGSTU1）被确定为番茄抗白粉病病原体抗性反应的关键基因。在本研究中，将ShGSTU1α克隆到质粒pET-28a中，使其在大肠埃希氏大肠杆菌异丙基-β-D-1-硫代半乳糖吡喃糖苷（IPTG）中有效表达，并用Ni2 +亲和层析纯化并进行生化鉴定。结果表明重组ShGSTU1在？E中表达的最佳条件。大肠杆菌在37°C和1 hM浓度的IPTG诱导下生长4小时。从1 g湿细菌中回收到约18.93 mg ShGSTU1。重组ShGSTU1表现出酶活性，比活性为0.625U / mg。这些结果可能为以后研究ShGSTU1在番茄抗白粉病机理中的研究提供重要依据。

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《African Journal of Biotechnology》 |2012年第23期|共7页
作者
Dongli Pei; Jinping Ding; Zilei Duan; Meng Li; Yiyang Feng; Chengwei Li;
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中图分类生物工程学（生物技术）;
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Tomatoglutathione S-transferaseexpressionpurificationenzyme activity;

机译：番茄谷胱甘肽S-转移酶表达纯化酶活性;

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1. Expression of lipase-solubilized bovine liver microsomal cytochrome b(5) in Escherichia coli as a glutathione S-transferase fusion protein (GST-cyt b(5)) [J] . Lin YW, Zhao DX, Wang ZH, Protein Expression and Purification . 2006,第2期

机译：脂酶溶解的牛肝微粒体细胞色素b（5）作为谷胱甘肽S-转移酶融合蛋白（GST-cyt b（5））在大肠杆菌中的表达
2. Increased solubility of glutathione S-transferase p16 (GST-p16) fusionprotein by co-expression of chaperones groES and groEL in Escherichia coli [J] . Luo ZH., Hua ZC. Biochemistry and Molecular Biology International . 1998,第3期

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3. Molecular cloning and characterization of omega class glutathione S-transferase (GST-O) from the polychaete Neanthes succinea: Biochemical comparison with theta class glutathione S-transferase (GST-T) [J] . Jae-Sung Rhee, Young-Mi Lee, Dae-Sik Hwang, Comparative biochemistry and physiology. Toxicology & pharmacology: CBP . 2007,第4期

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4. Expression of a fusion protein of human proinsulin with glutathione-S-transferase in Escherichia coli [C] . Zhixin Di, Jianzhong Ma, Yonggang Wang International Conference on Applied Sciences, Engineering and Technology . 2014

机译：人咪素素与谷胱甘肽-S-转移酶在大肠杆菌中的表达
5. Analysis of glutathione S-transferase (GST) gene expression in transgenic plants and its effects on environmental stress. [D] . Kang, Won Hee. 1998

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6. Cloning expression and biochemical characterization of one Epsilon-class (GST-3) and ten Delta-class (GST-1) glutathione S-transferases from Drosophila melanogaster and identification of additional nine members of the Epsilon class. [O] . Rafał Sawicki, Sharda P Singh, Ashis K Mondal, 2003

机译：果蝇果蝇的一种Epsilon类（GST-3）和十种Delta类（GST-1）谷胱甘肽S-转移酶的克隆表达和生化特征以及鉴定Epsilon类的另外九个成员。
7. Cloning, expression and biochemical characterization of one Epsilon-class (GST-3) and ten Delta-class (GST-1) glutathione S-transferases from Drosophila melanogaster, and identification of additional nine members of the Epsilon class. [O] . Sawicki, Rafał, Singh, Sharda P, Mondal, Ashis K, 2003

机译：果蝇果蝇的一种Epsilon类（GST-3）和十种Delta类（GST-1）谷胱甘肽S-转移酶的克隆，表达和生化特征，以及鉴定Epsilon类的另外九个成员。

Cloning and expression of a tomato glutathione S- transferase (GST) in Escherichia coli

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