Identification, Characterization, and Molecular Application of a Virulence-Associated Autotransporter from a Pathogenic Pseudomonas fluorescens Strain
文献类型：期刊论文
| 作者 | Hu, Yong-hua1,2; Liu, Chun-sheng1,2; Hou, Jin-hui3; Sun, Li1; Sun, L, Chinese Acad Sci, Inst Oceanol, 7 Nanhai Rd, Qingdao 266071, Peoples R China |
| 刊名 | APPLIED AND ENVIRONMENTAL MICROBIOLOGY
 |
| 出版日期 | 2009-07-01 |
| 卷号 | 75期号:13页码:4333-4340 |
| 关键词 | Periplasmic Chaperone Skp Cell-surface Presentation Gram-negative Bacteria v Protein Secretion Escherichia-coli Trimeric Autotransporter Serine-protease Yersinia-enterocolitica Recombinant Proteins Translocator Domain |
| ISSN号 | 0099-2240 |
| DOI | 10.1128/AEM.00159-09 |
| 文献子类 | Article |
| 英文摘要 | A gene, pfa1, encoding an autotransporter was cloned from a pathogenic Pseudomonas fluorescens strain, TSS, isolated from diseased fish. The expression of pfa1 is enhanced during infection and is regulated by growth phase and growth conditions. Mutation of pfa1 significantly attenuates the overall bacterial virulence of TSS and impairs the abilities of TSS in biofilm production, interaction with host cells, modulation of host immune responses, and dissemination in host blood. The putative protein encoded by pfa1 is 1,242 amino acids in length and characterized by the presence of three functional domains that are typical for autotransporters. The passenger domain of PfaI contains a putative serine protease (Pap) that exhibits apparent proteolytic activity when expressed in and purified from Escherichia coli as a recombinant protein. Consistent with the important role played by PfaI in bacterial virulence, purified recombinant Pap has a profound cytotoxic effect on cultured fish cells. Enzymatic analysis showed that recombinant Pap is relatively heat stable and has an optimal temperature and pH of 50 degrees C and pH 8.0. The domains of PfaI that are essential to autotransporting activity were localized, and on the basis of this, a PfaI-based autodisplay system (named AT1) was engineered to facilitate the insertion and transport of heterologous proteins. When expressed in E. coli, AT1 was able to deliver an integrated Edwardsiella tarda immunogen (Et18) onto the surface of bacterial cells. Compared to purified recombinant Et18, Et18 displayed by E. coli via AT1 induced significantly enhanced immunoprotection.; A gene, pfa1, encoding an autotransporter was cloned from a pathogenic Pseudomonas fluorescens strain, TSS, isolated from diseased fish. The expression of pfa1 is enhanced during infection and is regulated by growth phase and growth conditions. Mutation of pfa1 significantly attenuates the overall bacterial virulence of TSS and impairs the abilities of TSS in biofilm production, interaction with host cells, modulation of host immune responses, and dissemination in host blood. The putative protein encoded by pfa1 is 1,242 amino acids in length and characterized by the presence of three functional domains that are typical for autotransporters. The passenger domain of PfaI contains a putative serine protease (Pap) that exhibits apparent proteolytic activity when expressed in and purified from Escherichia coli as a recombinant protein. Consistent with the important role played by PfaI in bacterial virulence, purified recombinant Pap has a profound cytotoxic effect on cultured fish cells. Enzymatic analysis showed that recombinant Pap is relatively heat stable and has an optimal temperature and pH of 50 degrees C and pH 8.0. The domains of PfaI that are essential to autotransporting activity were localized, and on the basis of this, a PfaI-based autodisplay system (named AT1) was engineered to facilitate the insertion and transport of heterologous proteins. When expressed in E. coli, AT1 was able to deliver an integrated Edwardsiella tarda immunogen (Et18) onto the surface of bacterial cells. Compared to purified recombinant Et18, Et18 displayed by E. coli via AT1 induced significantly enhanced immunoprotection. |
| 学科主题 | Biotechnology & Applied Microbiology ; Microbiology |
| URL标识 | 查看原文 |
| 语种 | 英语 |
| WOS记录号 | WOS:000267373000013 |
| 公开日期 | 2010-11-18 |
| 源URL | [http://ir.qdio.ac.cn/handle/337002/1606]  |
| 专题 | 海洋研究所_实验海洋生物学重点实验室 |
| 通讯作者 | Sun, L, Chinese Acad Sci, Inst Oceanol, 7 Nanhai Rd, Qingdao 266071, Peoples R China |
| 作者单位 | 1.Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China 2.Chinese Acad Sci, Grad Univ, Beijing, Peoples R China 3.Xuzhou Inst Technol, Xuzhou, Peoples R China |
| 推荐引用方式 GB/T 7714 | Hu, Yong-hua,Liu, Chun-sheng,Hou, Jin-hui,et al. Identification, Characterization, and Molecular Application of a Virulence-Associated Autotransporter from a Pathogenic Pseudomonas fluorescens Strain[J]. APPLIED AND ENVIRONMENTAL MICROBIOLOGY,2009,75(13):4333-4340. |
| APA | Hu, Yong-hua,Liu, Chun-sheng,Hou, Jin-hui,Sun, Li,&Sun, L, Chinese Acad Sci, Inst Oceanol, 7 Nanhai Rd, Qingdao 266071, Peoples R China.(2009).Identification, Characterization, and Molecular Application of a Virulence-Associated Autotransporter from a Pathogenic Pseudomonas fluorescens Strain.APPLIED AND ENVIRONMENTAL MICROBIOLOGY,75(13),4333-4340. |
| MLA | Hu, Yong-hua,et al."Identification, Characterization, and Molecular Application of a Virulence-Associated Autotransporter from a Pathogenic Pseudomonas fluorescens Strain".APPLIED AND ENVIRONMENTAL MICROBIOLOGY 75.13(2009):4333-4340. |
入库方式： OAI收割
来源：海洋研究所
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