Yamamoto Kohji, Nagaoka Sumiharu, Banno Yutaka, Aso Yoichi
Institute of Genetic Resources, Faculty of Agriculture, Kyushu University Graduate School, Higashi-ku, Fukuoka 812-8581, Japan.
Comp Biochem Physiol C Toxicol Pharmacol. 2009 May;149(4):461-7. doi: 10.1016/j.cbpc.2008.10.108. Epub 2008 Nov 3.
A cDNA encoding an omega-class glutathione S-transferase of the silkmoth, Bombyx mori (bmGSTO), was cloned by reverse transcriptase-polymerase chain reaction. The resulting clone was sequenced and deduced for amino acid sequence, which revealed 40, 40, and 39% identities to omega-class GSTs from human, pig, and mouse, respectively. A recombinant protein (rbmGSTO) was functionally overexpressed in Escherichia coli cells in a soluble form and purified to homogeneity. rbmGSTO was able to catalyze the biotranslation of glutathione with 1-chloro-2,4-dinitrobenzene, a model substrate for GST, as well as with 4-hydroxynonenal, a product of lipid peroxidation. This enzyme was shown to have high affinity for organophosphorus insecticide and was present abundantly in silkmoth strain exhibiting fenitrothion resistance. These results indicate that bmGSTO could be involved in the increase in level of insecticide resistance for lepidopteran insects.
通过逆转录聚合酶链反应克隆了家蚕（Bombyx mori）编码ω-类谷胱甘肽S-转移酶（bmGSTO）的cDNA。对所得克隆进行测序并推导氨基酸序列，结果显示其与人、猪和小鼠的ω-类谷胱甘肽S-转移酶的同源性分别为40%、40%和39%。重组蛋白（rbmGSTO）在大肠杆菌细胞中以可溶形式功能性过表达并纯化至同质。rbmGSTO能够催化谷胱甘肽与1-氯-2,4-二硝基苯（谷胱甘肽S-转移酶的模型底物）以及脂质过氧化产物4-羟基壬烯醛的生物转化。该酶对有机磷杀虫剂具有高亲和力，并且大量存在于对杀螟硫磷具有抗性的家蚕品系中。这些结果表明bmGSTO可能参与鳞翅目昆虫抗杀虫剂水平的提高。
