Caggese C, Ragone G, Perrini B, Moschetti R, De Pinto V, Caizzi R, Barsanti P
Istituto di Genetica, Università di Bari, Italy.
Mol Gen Genet. 1999 Feb;261(1):64-70. doi: 10.1007/s004380050942.
As a first step towards using cross-species comparison to complete the inventory of the nuclear genes that encode mitochondrial polypeptides, and ultimately to understand their function through systematic molecular and genetic analysis in a model organism of choice, we report here the characterization of 41 Drosophila melanogaster cDNAs. These cDNAs were isolated by screening an ovarian expression library with antibodies against mitochondrial proteins and identify 17 novel Drosophila genes. The deduced amino acid sequences encoded by the majority of these cDNAs turned out to show significant homology to mitochondrial proteins previously identified in other species. Among others, ORFs putatively encoding six different subunits of ATP synthase and three NADH:ubiquinone reductase subunits were detected. By in situ hybridization, all cDNAs were mapped to single bands on polytene chromosomes, thus identifying candidate Drosophila genes required for mitochondrial biogenesis and maintenance. A search of the Human Gene Index database made it possible in most cases to align the entire Drosophila coding sequence with a human consensus sequence, suggesting that the cDNAs originate from insect counterparts of expressed mammalian genes. Our experimental strategy represents an efficient approach to the identification and interspecies comparison of genes encoding products targeted to the mitochondrion.
作为利用跨物种比较来完成编码线粒体多肽的核基因清单,并最终通过在选定的模式生物中进行系统的分子和遗传分析来了解其功能的第一步,我们在此报告41个黑腹果蝇cDNA的特征。这些cDNA是通过用抗线粒体蛋白抗体筛选卵巢表达文库分离得到的,鉴定出17个新的果蝇基因。结果表明,这些cDNA中的大多数所编码的推导氨基酸序列与先前在其他物种中鉴定出的线粒体蛋白具有显著同源性。其中,检测到推定编码ATP合酶六个不同亚基和三个NADH:泛醌还原酶亚基的开放阅读框。通过原位杂交,所有cDNA都定位到多线染色体上的单条带,从而鉴定出线粒体生物发生和维持所需的候选果蝇基因。对人类基因索引数据库的搜索使得在大多数情况下能够将整个果蝇编码序列与人类共有序列进行比对,这表明这些cDNA源自表达的哺乳动物基因的昆虫对应物。我们的实验策略是一种鉴定和跨物种比较编码靶向线粒体产物的基因的有效方法。