Identification and Functional Analysis of Novel Virulence Genes in Clostridium perfringens.

产气荚膜梭菌新毒力基因的鉴定和功能分析。

• 批准号: 09670276
• 负责人: SHIMIZU Tohru
• 金额: $ 1.98万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670276/
• 关键词: Clostridium perfringens; differential display; virulence; two-component system; genetic regulation; ウェルシュ菌; 二成分制御系; 転写調節; 病原性

Novel genes which are regulated in Clostridium perfringens by the two-component regulatory system, VirR/VirS, were identified using a differential display method. A plasmid library was constructed from C.perfringens chromosomal DNA and total of 1,200 independent plasmids was hybridized with cDNA probes prepared from total RNA of wild type strain 13 and its virR-mutant derivative TS133. Three clones were newly identified as being regulated by VirR/VirS, two of which were positively- and the other negatively-regulated. The fragment on which the VirR/VirS-regulated genes were located was identified by hybridizing the plasmid DNA with cDNA probes, and the nucleotide sequence of each fragment was determined. Several open reading frames were found and some of their deduced amino acid sequences showed significant similarities with protein-tyrosine phosphatase, 2', 3'-cyclic nucleotide 2'-phosphodiesterase, and cysteine synthase. The results suggested that the global regulatory system VirR/VirS should regulate various genes, other than toxin genes, both positively and negatively, and the newly identified genes were suspected to encode novel virulence factors in C.perfringens.

用差异显示方法鉴定了产气荚膜梭菌中受VirR/VIRS双组分调控系统调控的新基因。以产气荚膜梭菌染色体DNA为模板构建了质粒库，将1200个独立的质粒与野生型13菌株及其病毒突变体TS133的总RNA制备的cDNADNA探针杂交。新发现的三个克隆受Virr/VIRS的调控，其中两个是正调控的，另一个是负调控的。将载体DNA与cDNA探针杂交，确定病毒/VIRS调控基因所在的片段，并测定各片段的核苷酸序列。发现了几个开放阅读框，其中一些推导的氨基酸序列与蛋白质酪氨酸磷酸酶、2‘，3’-环核苷酸2‘-磷酸二酯酶和半胱氨酸合成酶有显著的相似性。结果表明，除毒素基因外，全球调控系统VirR/VIRS应对多种基因进行正向和负向调控，新发现的基因可能编码产气荚膜梭菌新的毒力因子。

项目成果

Ba-Thein, W.et al.: "Genomic diversity in the pfoA region of theta-toxin deficient strains of Clostridium perfringens." Microbiol.Immunol.41. 629-631 (1997)

Ba-Thein, W.等人：“产气荚膜梭菌 theta 毒素缺陷菌株 pfoA 区域的基因组多样性。”

William Ba-Thein: "Genomic diversity in the pfoA regron ot thera-torin-deficient stuains ot clostriclium penfungens" Microbool. Journol. 41. 629-631 (1997)

William Ba-Thein：“pfoA regron ot thera-torin-deficient stuains ot clostriclium penfungens 的基因组多样性”Microbool。

Kaori Ohtani: "Collagenase gene (colA) is located in the 3′-flanking rogron of the perfringolysm O (pfoA) locus in Clostridium perfringens." FEMS Mierobiology Letters. 146. 155-159 (1997)

Kaori Ohtani：“胶原酶基因 (colA) 位于产气荚膜梭菌产气荚膜溶解 O (pfoA) 基因座的 3 侧翼。”FEMS Mierbiology Letters 146. 155-159 (1997)。

Kaori Ohtani: "Collagenase gene (colA) is lucated in the 3'-flanking region of the pertringolysin O (pfoA) locus in Clostridium perfringens." FEMS Microbiology Letters. 146. 155-159 (1997)

Kaori Ohtani：“胶原酶基因 (colA) 位于产气荚膜梭菌中产气荚膜杆菌溶血素 O (pfoA) 位点的 3 侧翼区域。”

清水 徹: "ウェルシュ菌における毒素産生調節機構の解析" 日本細菌学雑誌. 52. 659-670 (1997)

Toru Shimizu：“产气荚膜梭菌毒素产生的调节机制分析”日本细菌学杂志 52. 659-670 (1997)。