Braun H P
Institut für Angewandte Genetik, Universität Hannover, Germany.
Mol Biol Rep. 1996;23(2):71-7. doi: 10.1007/BF00424432.
Large-scale random cDNA sequencing projects have been started for several organisms and are a valuable tool for the analysis of quantitative and qualitative aspects of gene expression. However, the reliability of the obtained data is limited as most of the clones are only partially analysed on one strand. As a consequence the sequence entries derived from random cDNA sequencing projects usually comprise incomplete open reading frames. They nevertheless define complete and reliable coding sequences, if two prerequisites are fulfilled: (i) the clones encode very small proteins, and (ii) the clones have a high frequency in the cDNA-banks. The present study describes the use of cDNA databases for the identification of homologues of three low-molecular-weight subunits of the mitochondrial bc1 complex, termed the QCR6, QCR9 and QCR10 proteins. These polypeptides are only characterized for a small number of organisms, have a scarcely defined function and exhibit a low degree of structural conservation if compared between different species. Several clones were identified for each polypeptide by searches with TBLASTN using the known sequences as probes. Most of the database entries contain complete open reading frames and sequencing queries could be excluded due to the abundancy of the clones. Multiple sequence alignments are presented for all three polypeptides and consensus sequences are given which may provide a basis for the investigation of the proteins by site-directed mutagenesis.
已经针对几种生物启动了大规模随机cDNA测序项目,这些项目是分析基因表达的定量和定性方面的宝贵工具。然而,由于大多数克隆仅在一条链上进行了部分分析,因此所获得数据的可靠性受到限制。结果,来自随机cDNA测序项目的序列条目通常包含不完整的开放阅读框。然而,如果满足两个先决条件,它们就能定义完整且可靠的编码序列:(i)克隆编码非常小的蛋白质,以及(ii)克隆在cDNA文库中的频率很高。本研究描述了利用cDNA数据库鉴定线粒体bc1复合物的三个低分子量亚基的同源物,即QCR6、QCR9和QCR10蛋白。这些多肽仅在少数生物中得到表征,功能定义不明确,并且在不同物种之间比较时结构保守程度较低。使用已知序列作为探针,通过TBLASTN搜索为每个多肽鉴定了几个克隆。大多数数据库条目包含完整的开放阅读框,并且由于克隆数量丰富,可以排除测序查询。给出了所有三种多肽的多序列比对,并给出了共有序列,这可能为通过定点诱变研究这些蛋白质提供基础。