Maneewannakul S, Kathir P, Moore D, Le L A, Wu J H, Ippen-Ihler K
Department of Medical Microbiology and Immunology, Texas A & M University, College Station 77843.
J Bacteriol. 1987 Nov;169(11):5119-24. doi: 10.1128/jb.169.11.5119-5124.1987.
As part of an analysis of the conjugative transfer genes associated with the expression of F pili by plasmid F, we have investigated the physical location of the traC and traW genes. We found that plasmid clones carrying a 2.95-kilobase EcoRI-EcoRV F transfer operon fragment were able to complement transfer of F lac traC mutants and expressed an approximately 92,000-dalton product that comigrates with TraC. We also found that traW-complementing activity was expressed from plasmids carrying a 900-base-pair SmaI-HincII fragment. The traW product was identified as an approximately 23,000-dalton protein. The two different F DNA fragments that expressed traC and traW activities do not overlap. Our data indicate that the traC gene is located in a more-tra operon promoter-proximal position than suggested on earlier maps and that traW is distal to traC. These results resolve a long-standing question concerning the relationship of traW to traC. The clones we have constructed are expected to be useful in elucidating the role of proteins TraC and TraW in F-pilus assembly.
作为对与质粒F表达F菌毛相关的接合转移基因分析的一部分,我们研究了traC和traW基因的物理位置。我们发现,携带2.95千碱基EcoRI - EcoRV F转移操纵子片段的质粒克隆能够互补F lac traC突变体的转移,并表达一种与TraC共迁移的约92,000道尔顿的产物。我们还发现,携带900碱基对SmaI - HincII片段的质粒表达traW互补活性。traW产物被鉴定为一种约23,000道尔顿的蛋白质。表达traC和traW活性的两个不同F DNA片段不重叠。我们的数据表明,traC基因位于比早期图谱所显示的更靠近tra操纵子启动子的位置,并且traW位于traC的远端。这些结果解决了一个长期存在的关于traW与traC关系的问题。我们构建的克隆有望用于阐明蛋白质TraC和TraW在F菌毛组装中的作用。