Richter S, Hess W R, Krause M, Messer W
Max-Planck-Institut für molekulare Genetik, Berlin, Germany.
Mol Gen Genet. 1998 Mar;257(5):534-41. doi: 10.1007/s004380050679.
In order to study DNA replication control elements in cyanobacteria we cloned and sequenced the dnaA gene from the marine cyanobacterium Prochlorococcus marinus. The dnaA gene is ubiquitous among bacteria and encodes the DNA replication initiation factor DnaA. The deduced amino acid sequence of the P. marinus DnaA protein shows highest similarity to the DnaA protein from the freshwater cyanobacterium Synechocystis sp. PCC6803. Using a solid-phase DNA binding assay we demonstrated that both cyanobacterial DnaA proteins specifically recognize chromosomal origins, oriC, of Escherichia coli and Bacillus subtilis in vitro. The genetic environment of dnaA is not conserved between the two cyanobacteria. Upstream of the P. marinus dnaA gene we identified a gene encoding a putative ATP-binding cassette (ABC) transport protein. The gor gene encoding glutathione reductase lies downstream of dnaA. Comparison of the genetic structure of dnaA regions from 15 representative bacteria shows that the pattern of genes flanking dnaA is not universally conserved among them.
为了研究蓝藻中的DNA复制控制元件,我们从海洋蓝藻聚球藻属的海聚球藻中克隆并测序了dnaA基因。dnaA基因在细菌中普遍存在,编码DNA复制起始因子DnaA。海聚球藻DnaA蛋白的推导氨基酸序列与淡水蓝藻集胞藻属的PCC6803的DnaA蛋白具有最高的相似性。使用固相DNA结合试验,我们证明两种蓝藻DnaA蛋白在体外都能特异性识别大肠杆菌和枯草芽孢杆菌的染色体起源oriC。两种蓝藻之间dnaA的遗传环境并不保守。在海聚球藻dnaA基因的上游,我们鉴定出一个编码假定ATP结合盒(ABC)转运蛋白的基因。编码谷胱甘肽还原酶的gor基因位于dnaA的下游。对15种代表性细菌的dnaA区域遗传结构的比较表明,dnaA侧翼基因的模式在它们之间并非普遍保守。