Völkl P, Markiewicz P, Baikalov C, Fitz-Gibbon S, Stetter K O, Miller J H
Department of Microbiology and Molecular Genetics, University of California, Los Angeles 90024, USA.
Nucleic Acids Res. 1996 Nov 15;24(22):4373-8. doi: 10.1093/nar/24.22.4373.
The hyperthermophilic archaeum, Pyrobaculum aerophilum, grows optimally at 100 degrees C with a doubling time of 180 min. It is a member of the phylogenetically ancient Thermoproteales order, but differs significantly from all other members by its facultatively aerobic metabolism. Due to its simple cultivation requirements and its nearly 100% plating efficiency, it was chosen as a model organism for studying the genome organization of hyperthermophilic ancient archaea. By a G+C content of the DNA of 52 mol%, sequence analysis was easily possible. At least some of the mRNA of P. aerophilum carried poly-A tails facilitating the construction of a cDNA library. 245 sequence tags of a poly-A primed cDNA library and 55 sequence tags from a 1-2 kb Sau3AI-fragment containing genomic library were analyzed and the corresponding amino acid sequences compared with protein sequences from databases. Fourteen percent of the cDNA and >9% of genomic DNA sequence tags revealed significant similarities to proteins in the databases. Matches were obtained to proteins from archaeal, bacterial and eukaryal sources. Some sequences showed greatest similarity to eukaryal rather than to bacterial versions of proteins, other matches were found to proteins which had previously only been found in eukaryotes.
嗜热古菌嗜气栖热袍菌在100℃时生长最佳，倍增时间为180分钟。它是系统发育上古老的栖热袍菌目的成员，但因其兼性有氧代谢而与所有其他成员有显著差异。由于其简单的培养要求和近100%的平板接种效率，它被选为研究嗜热古老古菌基因组组织的模式生物。由于其DNA的G+C含量为52mol%，因此很容易进行序列分析。嗜气栖热袍菌的至少一些mRNA带有聚腺苷酸尾，这便于构建cDNA文库。分析了聚腺苷酸引发的cDNA文库的245个序列标签和来自包含基因组文库的1-2kb Sau3AI片段的55个序列标签，并将相应的氨基酸序列与数据库中的蛋白质序列进行比较。14%的cDNA和>9%的基因组DNA序列标签与数据库中的蛋白质有显著相似性。与古菌、细菌和真核生物来源中的蛋白质进行了匹配。一些序列与真核生物版本的蛋白质而不是细菌版本的蛋白质显示出最大的相似性，还发现其他匹配的蛋白质以前仅在真核生物中发现。
