Oppentocht Jantien E, van Delden Wilke, van de Zande Louis
Population Genetics, Center for Ecological and Evolutionary Studies, Biological Center, University of Groningen, The Netherlands.
Mol Biol Evol. 2002 Jul;19(7):1026-40. doi: 10.1093/oxfordjournals.molbev.a004162.
The nucleotide sequences of the Adh and Adhr genes of Drosophila kuntzei were derived from combined overlapping sequences of clones isolated from a genomic library and from cloned PCR and inverse-PCR fragments. Only a proximal promoter was detected upstream of the Adh gene, indicating that D. kuntzei Adh is regulated by a one-promoter system. Further upstream of the Adh structural gene, an adult enhancer region (AAE) was found that contains most of the regulatory sequences described for AAEs of other Drosophila species. Analysis of the ADH protein showed an amino acid change from valine to threonine in the active site at position 189 which is also found in D. funebris but is otherwise unique among Drosophila. This difference alone may be responsible for the very low ADH activity found in this species and may cause a difference in substrate usage pattern. Codon bias in Adh and Adhr was comparable and found to be very low compared with other species. Phylogenetic analysis showed that D. kuntzei is closest related to D. funebris and D. immigrans. The time of divergence between D. kuntzei and D. funebris was estimated to be 14.2-20.2 Myr and that between D. kuntzei-D. funebris and D. immigrans to be 30.8-44.0 Myr. An analysis of the genetic variation in the Adh gene and upstream sequences of four European strains showed that this gene was highly variable. Overall nucleotide diversity (pi) was 0.0139, which is two times higher than that in D. melanogaster.
昆氏果蝇（Drosophila kuntzei）的乙醇脱氢酶（Adh）基因和乙醇脱氢酶相关基因（Adhr）的核苷酸序列，源自从基因组文库中分离出的克隆的重叠序列组合，以及克隆的聚合酶链式反应（PCR）片段和反向PCR片段。在Adh基因上游仅检测到一个近端启动子，这表明昆氏果蝇的Adh受单启动子系统调控。在Adh结构基因的更上游，发现了一个成虫增强子区域（AAE），其中包含了其他果蝇物种AAE中描述的大部分调控序列。对ADH蛋白的分析表明，在第189位的活性位点上，氨基酸从缬氨酸变为苏氨酸，这种变化在黑腹果蝇（D. funebris）中也有发现，但在果蝇中其他物种中是独特的。仅这一差异可能就导致了该物种中发现的极低的ADH活性，并可能导致底物使用模式的差异。Adh和Adhr中的密码子偏好性相当，与其他物种相比非常低。系统发育分析表明，昆氏果蝇与黑腹果蝇和迁入果蝇（D. immigrans）关系最为密切。昆氏果蝇和黑腹果蝇之间的分歧时间估计为1420 - 2020万年前，昆氏果蝇 - 黑腹果蝇与迁入果蝇之间的分歧时间为3080 - 4400万年前。对四个欧洲菌株的Adh基因及其上游序列的遗传变异分析表明，该基因具有高度变异性。总体核苷酸多样性（pi）为0.0139，是黑腹果蝇的两倍。
