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| Expression and Characterization of a Sigma-Class Glutathione S-transferase of the Oriental Migratory Locust, Locusta migratoria manilensis (Meyen) |
| JIA Miao, QIN Guo-hua, LIU Ting, ZHANG Jian-zhen, ZHANG Xue-yao, ZHU Kun-yan, GUO Yaping, MA En-bo |
1.Research Institute of Applied Biology, Shanxi University
2.College of Environmental Science and Resource, Shanxi University
3.Department of Entomology, 123 Waters Hall, Kansas State University
4.School of Life Science, Shanxi University |
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摘要 A cDNA encoding a sigma-class glutathione S-transferase of the locust, Locusta migratoria manilensis (LmGSTs1), was cloned by reverse transcriptase-polymerase chain reaction. The 830 bp-long cDNA encoded a 615 bp open reading frame (204 amino acid polypeptide), which exhibited the structural motif and domain organization characteristic of GST sigmaclass. It revealed 59, 57, 57, and 56% identities to sigma-class GSTs from Blattella germanica, Gryllotalpa orientalis, Nasonia vitripennis, and Pediculus humanus corporis, respectively. A recombinant protein (LmGSTs1) was functionally expressed in Escherichia coli cells in a soluble form and purified to homogeneity. LmGSTs1 was able to catalyze the biotranslation of glutathione with 1-chloro-2,4-dinitrobenzene, a model substrate for GSTs, as well as with p-nitro-benzyl chloride. Its optimal activity was observed at pH 8.0 and at 30°C. Incubation for 30 min at temperatures below 50°C scarcely affected the activity. The I50 of reactive blue (RB) was 18.5 μmol L-1. In the presence of 0.05 mmol L-1 ethacrynic acid (ECA), LmGSTs1 showed (81±3)% of the original activities.
Abstract A cDNA encoding a sigma-class glutathione S-transferase of the locust, Locusta migratoria manilensis (LmGSTs1), was cloned by reverse transcriptase-polymerase chain reaction. The 830 bp-long cDNA encoded a 615 bp open reading frame (204 amino acid polypeptide), which exhibited the structural motif and domain organization characteristic of GST sigmaclass. It revealed 59, 57, 57, and 56% identities to sigma-class GSTs from Blattella germanica, Gryllotalpa orientalis, Nasonia vitripennis, and Pediculus humanus corporis, respectively. A recombinant protein (LmGSTs1) was functionally expressed in Escherichia coli cells in a soluble form and purified to homogeneity. LmGSTs1 was able to catalyze the biotranslation of glutathione with 1-chloro-2,4-dinitrobenzene, a model substrate for GSTs, as well as with p-nitro-benzyl chloride. Its optimal activity was observed at pH 8.0 and at 30°C. Incubation for 30 min at temperatures below 50°C scarcely affected the activity. The I50 of reactive blue (RB) was 18.5 μmol L-1. In the presence of 0.05 mmol L-1 ethacrynic acid (ECA), LmGSTs1 showed (81±3)% of the original activities.
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Received: 06 September 2010
Accepted:
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| Fund: Our research was supported by the Funds for International Cooperation and Exchange of the National Natural Science Foundation of China (30810103907), the National Natural Science Foundation of China (30870302), the Public Welfare Fund for Agriculture, Ministry of Agriculture, China (200903021), and the China Postdoctoral Science Foundation (Special Program, 201003656; Regular Program, 20090451359). |
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Corresponding Authors:
Correspondence MA En-bo, Professor, Tel/Fax: +86-351-7018871, E-mail: maenbo2003@sxu.edu.cn
E-mail: maenbo2003@sxu.edu.cn
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| About author: JIA Miao, MSc candidate, E-mail: jiamiao00013@163.com |
Cite this article:
JIA Miao, QIN Guo-hua, LIU Ting, ZHANG Jian-zhen, ZHANG Xue-yao, ZHU Kun-yan, GUO Yaping, MA En-bo.
2011.
Expression and Characterization of a Sigma-Class Glutathione S-transferase of the Oriental Migratory Locust, Locusta migratoria manilensis (Meyen) . Journal of Integrative Agriculture, 10(10): 1570-1576.
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