Syvanen M, Zhou Z H, Wang J Y
School of Medicine, University of California Davis 95616.
Mol Gen Genet. 1994 Oct 17;245(1):25-31. doi: 10.1007/BF00279747.
Three new glutathione transferase (GST) genes from the housefly Musca domestica are described. These genes, identified as MdGST-2, -3, and -4, were from cDNA clones obtained from a cDNA bank in phage lambda. The bank was prepared using poly(A)+ RNA from a housefly that is highly resistant to organophosphate insecticides because of enhanced expression of multiple members of the glutathione transferase gene family. The DNA sequence of each is reported and has a complete open reading frame that specified an amino acid sequence similar to other dipteran glutathione transferases. Based on phylogenetic analysis, we can conclude that the insect glutathione transferase gene family falls into two groups, each of which evolves at a different rate, presumably due to differences in functional constraints. We show that MdGST-1 (and their homologues from Drosophila and Lucilia) evolve at a significantly slower rate than the other members of the gene family. Each housefly GST cDNA was inserted into a bacterial plasmid expression system and a glutathione transferase activity was expressed in Escherichia coli. The transcription pattern of each of these glutathione transferases was examined in a variety of different housefly strains that are known to differ in their resistance to organophosphate insecticides due to different patterns of glutathione transferase expression. We found that the level of transcription for two of our clones was positively correlated with the level of organophosphate resistance.
本文描述了家蝇(Musca domestica)中的三个新的谷胱甘肽S-转移酶(GST)基因。这些基因被鉴定为MdGST-2、-3和-4,来自从噬菌体λ cDNA文库中获得的cDNA克隆。该文库是使用来自一只对有机磷杀虫剂具有高度抗性的家蝇的聚腺苷酸加尾RNA(poly(A)+ RNA)构建的,该家蝇由于谷胱甘肽S-转移酶基因家族多个成员的表达增强而具有抗性。报道了每个基因的DNA序列,它们都有一个完整的开放阅读框,其编码的氨基酸序列与其他双翅目谷胱甘肽S-转移酶相似。基于系统发育分析,我们可以得出结论,昆虫谷胱甘肽S-转移酶基因家族分为两组,每组以不同的速率进化,推测是由于功能限制的差异。我们发现MdGST-1(以及来自果蝇和绿蝇的同源物)的进化速率明显低于该基因家族的其他成员。将每个家蝇GST cDNA插入细菌质粒表达系统,并在大肠杆菌中表达谷胱甘肽S-转移酶活性。在多种已知因谷胱甘肽S-转移酶表达模式不同而对有机磷杀虫剂抗性不同的家蝇品系中,检测了这些谷胱甘肽S-转移酶各自的转录模式。我们发现,我们的两个克隆的转录水平与有机磷抗性水平呈正相关。