Lai C, McMahon R, Young C, Mackay T F, Langley C H
Center for Population Biology, University of California, Davis, California 95616, USA.
Genetics. 1998 Jun;149(2):1051-61. doi: 10.1093/genetics/149.2.1051.
The quemao (qm) locus of Drosophila melanogaster is characterized by a P-element-associated mutant lacking most of the large bristles on the thorax and by several EMS-induced recessive lethals. quemao was cloned using a transposon tagging strategy. P-element-mediated transformation demonstrated that the cloned qm DNA sequence (from the 65F cytological region) rescues the mutant phenotype. A 2.3-kb qm transcript was identified by Northern blot analysis by sequencing of the isolated qm cDNA clones and by 5' rapid amplification cDNA end (RACE). The predicted amino acid sequence (338 residues) of the coding region of the qm transcript shares 42, 31, 13, 20, and 12% identical amino acid sequences with the geranylgeranyl pyrophosphate synthase (GGPPS) of fungi, yeast, plants, archaebacteria, and eubacteria, respectively. It also contains five highly conserved domains common among all known isoprenyl pyrophosphate synthases. The P element associated with the original qm mutant is inserted in the 5' untranslated region of the transcript. An EMS-induced qm nonsense mutation at the 12th codon leads to recessive lethality at the first larval instar, indicating the essential role of qm in the isoprenoid biosynthesis of insects.
黑腹果蝇的quemao(qm)基因座的特征是一个与P因子相关的突变体,其胸部大部分大刚毛缺失,还有几个经乙基亚硝基脲(EMS)诱导产生的隐性致死突变。qm是采用转座子标签策略克隆得到的。P因子介导的转化实验表明,克隆得到的qm DNA序列(来自65F细胞学区域)能够拯救突变表型。通过Northern印迹分析、对分离得到的qm cDNA克隆进行测序以及5'端快速扩增cDNA末端(RACE)技术,鉴定出一个2.3 kb的qm转录本。qm转录本编码区预测的氨基酸序列(338个残基)与真菌、酵母、植物、古细菌和真细菌的牻牛儿基牻牛儿基焦磷酸合酶(GGPPS)的氨基酸序列分别有42%、31%、13%、20%和12%的相同性。它还包含所有已知异戊二烯基焦磷酸合酶共有的五个高度保守结构域。与原始qm突变体相关的P因子插入在转录本的5'非翻译区。在第12个密码子处经EMS诱导产生的qm无义突变导致一龄幼虫期隐性致死,这表明qm在昆虫类异戊二烯生物合成中起关键作用。