Chuang Yin-Ching, Wang Ke-Chuan, Chen Yi-Tseng, Yang Chia-Huei, Men Shang-Chin, Fan Chia-Chun, Chang Li-Huan, Yeh Kuang-Sheng
Department of Medical Research, Chi Mei Medical Center, 901 Chung Hwa Road, Yong Kang City, Tainan 710, Taiwan.
BMC Microbiol. 2008 Jul 25;8:126. doi: 10.1186/1471-2180-8-126.
Type 1 fimbriae are the most commonly found fimbrial appendages on the outer membrane of Salmonella enterica serotype Typhimurium. Previous investigations indicate that static broth culture favours S. Typhimurium to produce type 1 fimbriae, while non-fimbriate bacteria are obtained by growth on solid agar media. The phenotypic expression of type 1 fimbriae in S. Typhimurium is the result of the interaction and cooperation of several genes in the fim gene cluster. Other gene products that may also participate in the regulation of type 1 fimbrial expression remain uncharacterized.
In the present study, transposon insertion mutagenesis was performed on S. Typhimurium to generate a library to screen for those mutants that would exhibit different type 1 fimbrial phenotypes than the parental strain. Eight-two mutants were obtained from 7,239 clones screened using the yeast agglutination test. Forty-four mutants produced type 1 fimbriae on both solid agar and static broth media, while none of the other 38 mutants formed type 1 fimbriae in either culture condition. The flanking sequences of the transposons from 54 mutants were cloned and sequenced. These mutants can be classified according to the functions or putative functions of the open reading frames disrupted by the transposon. Our current results indicate that the genetic determinants such as those involved in the fimbrial biogenesis and regulation, global regulators, transporter proteins, prophage-derived proteins, and enzymes of different functions, to name a few, may play a role in the regulation of type 1 fimbrial expression in response to solid agar and static broth culture conditions. A complementation test revealed that transforming a recombinant plasmid possessing the coding sequence of a NAD(P)H-flavin reductase gene ubiB restored an ubiB mutant to exhibit the type 1 fimbrial phenotype as its parental strain.
Genetic determinants other than the fim genes may involve in the regulation of type 1 fimbrial expression in S. Typhimurium. How each gene product may influence type 1 fimbrial expression is an interesting research topic which warrants further investigation.
1型菌毛是鼠伤寒沙门氏菌血清型鼠伤寒菌株外膜上最常见的菌毛附属物。先前的研究表明,静态肉汤培养有利于鼠伤寒沙门氏菌产生1型菌毛,而在固体琼脂培养基上生长可获得无菌毛细菌。鼠伤寒沙门氏菌中1型菌毛的表型表达是fim基因簇中多个基因相互作用与协作的结果。其他可能也参与1型菌毛表达调控的基因产物仍未得到表征。
在本研究中,对鼠伤寒沙门氏菌进行转座子插入诱变以构建文库,筛选出那些与亲本菌株相比表现出不同1型菌毛表型的突变体。使用酵母凝集试验从7239个克隆中筛选出82个突变体。44个突变体在固体琼脂和静态肉汤培养基上均产生1型菌毛,而其他38个突变体在任何一种培养条件下均未形成1型菌毛。克隆并测序了54个突变体中转座子的侧翼序列。这些突变体可根据被转座子破坏的开放阅读框的功能或推定功能进行分类。我们目前的结果表明,诸如参与菌毛生物合成和调控、全局调控因子、转运蛋白、前噬菌体衍生蛋白以及不同功能的酶等遗传决定因素,可能在响应固体琼脂和静态肉汤培养条件时对1型菌毛表达的调控中发挥作用。互补试验表明,转化具有NAD(P)H-黄素还原酶基因ubiB编码序列的重组质粒可使ubiB突变体恢复为表现出与亲本菌株相同的1型菌毛表型。
除fim基因外,遗传决定因素可能参与鼠伤寒沙门氏菌中1型菌毛表达的调控。每个基因产物如何影响1型菌毛表达是一个有趣的研究课题,值得进一步研究。