Uemori T, Ishino Y, Toh H, Asada K, Kato I
Biotechnology Research Laboratories, Takara Shuzo Co., Ltd, Shiga, Japan.
Nucleic Acids Res. 1993 Jan 25;21(2):259-65. doi: 10.1093/nar/21.2.259.
We cloned the gene encoding the thermostable DNA polymerase from the archaeon Pyrococcus furiosus. The DNA fragment of 2785 base pair (bp) containing the structural gene for DNA polymerase was sequenced. DNA polymerase (Pfu polymerase), as deduced from the DNA sequence, consisted of 775 amino acids, had a molecular weight of 90, 109, and was structurally homologous to the alpha-like DNA polymerases (family B) represented by human DNA polymerase alpha and Escherichia coli DNA polymerase II. An unrooted phylogenetic tree of the alpha-like DNA polymerases based on the amino acid sequence alignment was constructed. Pfu polymerase, with two other archaeon polymerases, constitutes a group with some animal viruses. The transcription initiation sites of the pol gene were identified by analysis of in vivo transcripts of both from P. furiosus and E. coli, and the promoters were assigned upstream of the pol coding region. A typical promoter sequence for the archaeon was found at a reasonable distance from the transcription initiation site in P. furiosus.
我们从嗜热栖热球菌（Pyrococcus furiosus）中克隆了编码耐热DNA聚合酶的基因。对包含DNA聚合酶结构基因的2785个碱基对（bp）的DNA片段进行了测序。根据DNA序列推导，DNA聚合酶（Pfu聚合酶）由775个氨基酸组成，分子量为90109，在结构上与以人类DNA聚合酶α和大肠杆菌DNA聚合酶II为代表的α类DNA聚合酶（B家族）同源。基于氨基酸序列比对构建了α类DNA聚合酶的无根系统发育树。Pfu聚合酶与另外两种古菌聚合酶与一些动物病毒构成一个类群。通过对嗜热栖热球菌和大肠杆菌体内转录本的分析，确定了pol基因的转录起始位点，并在pol编码区上游确定了启动子。在嗜热栖热球菌中，在距转录起始位点合理距离处发现了典型的古菌启动子序列。
