琼氏黄杆菌（约翰逊氏噬纤维菌）滑行运动基因gldA的克隆与特性分析
Cloning and characterization of the Flavobacterium johnsoniae (Cytophaga johnsonae) gliding motility gene, gldA.
作者信息
Agarwal S, Hunnicutt D W, McBride M J
机构信息
Department of Biological Sciences, University of Wisconsin, P.O. Box 413, Milwaukee, WI 53201, USA.
出版信息
Proc Natl Acad Sci U S A. 1997 Oct 28;94(22):12139-44. doi: 10.1073/pnas.94.22.12139.
Abstract
The mechanism of bacterial gliding motility (active movement over surfaces without the aid of flagella) is not known. A large number of nonmotile mutants of the gliding bacterium Flavobacterium johnsoniae (Cytophaga johnsonae) have been previously isolated, and genetic techniques to analyze these mutants have recently been developed. We complemented a nonmotile mutant of F. johnsoniae (UW102-09) with a library of wild-type DNA by using the shuttle cosmid pCP17. The complementing plasmid (pCP100) contained an insert of 13 kbp, and restored motility to 4 of 61 independently isolated nonmotile mutants. A 1.3-kbp fragment that encompassed a single ORF, gldA, complemented all four mutants. Disruption of the chromosomal copy of gldA in wild-type F. johnsoniae UW101 eliminated gliding motility. The predicted protein produced by gldA has strong sequence similarity to ATP binding cassette transport proteins.
摘要
细菌滑行运动（在无鞭毛辅助的情况下在表面进行主动运动）的机制尚不清楚。先前已分离出大量滑行细菌约翰逊黄杆菌（约翰逊噬纤维菌）的非运动突变体，并且最近已开发出分析这些突变体的遗传技术。我们使用穿梭粘粒pCP17，用野生型DNA文库对约翰逊黄杆菌的一个非运动突变体（UW102 - 09）进行了互补。互补质粒（pCP100）含有一个13 kbp的插入片段，并使61个独立分离的非运动突变体中的4个恢复了运动能力。一个包含单个开放阅读框gldA的1.3 kbp片段，对所有四个突变体都起到了互补作用。在野生型约翰逊黄杆菌UW101中破坏gldA的染色体拷贝消除了滑行运动。gldA产生的预测蛋白与ATP结合盒转运蛋白具有很强的序列相似性。
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