Isolation and characterization of a cDNA clone coding for a glutathione S-transferase class delta enzyme from the biting midge Culicoides variipennis sonorensis Wirth and Jones
Isolation and characterization of a cDNA clone coding for a glutathione S-transferase class delta enzyme from the biting midge Culicoides variipennis sonorensis Wirth and Jones
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DOI：
10.1023/a:1026471830868
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发表时间：
2000-12-01
影响因子：
2.4
通讯作者：
Murphy, KE
中科院分区：
文献类型：
--
作者：
Abdallah, MA;Pollenz, RS;Murphy, KE
索诺库蠓 (Culicoides variipennis sonorensis) 是北美蓝舌病病毒的主要载体。谷胱甘肽 S-转移酶 (GST) 是催化亲核取代、将反应性亲脂性分子转化为可溶性缀合物的酶。 GST 活性的增加与杀虫剂抗药性的发展有关。这里描述的是编码 C. variipennis GST 的第一个 cDNA 的分离。该克隆由 720 个翻译碱基组成，编码 M-r 类似于 24,800 个由 219 个氨基酸组成的蛋白质。推导的氨基酸序列与双翅目蝇属的 Delta 类（以前称为 Theta）GST 相似（64%-74%）。果蝇、绿蝇和按蚊。该 cDNA 被亚克隆到 pET-11b 中，在伊壁鸠鲁大肠杆菌 BL21 (DE3) 中表达，对底物 1-氯-2,4-二硝基苯具有类似于 28,000 单位/毫克的比活性。
Culicoides variipennis sonorensis is the primary vector of bluetongue viruses in North America. Glutathione S-transferases (GSTs) are enzymes that catalyze nucleophilic substitutions, converting reactive lipophilic molecules into soluble conjugates. Increased GST activity is associated with development of insecticide resistance. Described here is the isolation of the first cDNA encoding a C. variipennis GST. The clone consists of 720 translated bases encoding a protein with a M-r of similar to 24,800 composed of 219 amino acids. The deduced amino acid sequence is similar (64%-74%) to class Delta (previously named Theta) GSTs from the dipteran genera Musca. Drosophila, Lucilia and Anopheles. The cDNA was subcloned into pET-11b, expressed in Epicurian coli BL21 (DE3) and has a specific activity of similar to 28,000 units/mg for the substrate 1-chloro-2,4-dinitrobenzene.