Finlay B B, Frost L S, Paranchych W
J Bacteriol. 1986 Oct;168(1):132-9. doi: 10.1128/jb.168.1.132-139.1986.
The complete nucleotide sequences of the ColB4-K98 (ColB4) plasmid transfer genes oriT, traM, and traY as well as the traY gene of R100-1 are presented and compared with the corresponding regions from the conjugative plasmids F, R1, and R100. The sequence encoding the oriT nick sites and surrounding inverted repeats identified in F was conserved in ColB4. The adenine-thymine-rich sequence following these nick sites was conserved in R1 and ColB4 but differed in F and R100, indicating that this region may serve as the recognition site for the traY protein. A series of direct repeats unique to the ColB4 plasmid was found in the region of dyad symmetry following this AT-rich region. This area also encodes 21-base-pair direct repeats which are homologous to those in F and R100. The traM gene product may bind in this region. Overlapping and following these repeats is the promoter(s) for the traM protein. The traM protein from ColB4 is similar to the equivalent products from F, R1, and R100. The traY protein from ColB4 is highly homologous to the R1 traY gene product, while the predicted R100-1 traY product differs at several positions. These differences presumably define the different alleles of traM and traY previously identified for IncF plasmids by genetic criteria. The translational start codons of the ColB4 and R100-1 traY genes are GUG and UUG, respectively, two examples of rare initiator codon usage.
本文展示了ColB4 - K98(ColB4)质粒转移基因oriT、traM和traY以及R100 - 1的traY基因的完整核苷酸序列,并将其与接合质粒F、R1和R100的相应区域进行了比较。在F中鉴定出的编码oriT切口位点及周围反向重复序列在ColB4中是保守的。这些切口位点之后富含腺嘌呤 - 胸腺嘧啶的序列在R1和ColB4中是保守的,但在F和R100中有所不同,这表明该区域可能是traY蛋白的识别位点。在这个富含AT的区域之后的二元对称区域中发现了一系列ColB4质粒特有的直接重复序列。该区域还编码与F和R100中的21个碱基对直接重复序列同源的序列。traM基因产物可能结合在该区域。在这些重复序列重叠并紧随其后的是traM蛋白的启动子。ColB4的traM蛋白与F、R1和R100的相应产物相似。ColB4的traY蛋白与R1的traY基因产物高度同源,而预测的R100 - 1的traY产物在几个位置有所不同。这些差异大概定义了先前通过遗传学标准为IncF质粒鉴定的traM和traY的不同等位基因。ColB4和R100 - 1的traY基因的翻译起始密码子分别是GUG和UUG,这是稀有起始密码子使用的两个例子。