• 主题
高级搜索  >
历史搜索 清空历史
首页> 美国政府科技报告 >Shewanella Federation: Functional Genomic Investigations of Dissimilatory Metal-Reducing Shewanella. Final Report
【24h】

Shewanella Federation: Functional Genomic Investigations of Dissimilatory Metal-Reducing Shewanella. Final Report

机译：shewanella联合会：异化金属减少希瓦氏菌的功能基因组研究。总结报告

获取原文
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Generation and validation of a Shewanella oneidensis MR-1 clone set for protein expression and phage display. An ORF clone set for S. oneidensis was created using the lambda recombinase system. ORFs within entry vectors in this system can be readily transferred into multiple destination vectors, making the clone set a useful resource for research groups studying this microorganism. To establish that the S. oneidensis clone set could be used for protein expression and functional studies, three sets of ORFs were examined for expression of His-tag proteins, expression of His/GST-tag proteins, or for effective display on phage. A total of 21 out of 30 (70%) predicted two-component transcriptional regulators from S. oneidensis were successfully expressed in the His-tag format. The use of the S. oneidensis clone set for functional studies was tested using a phage display system. The method involves the fusion of peptides or proteins to a coat protein of a bacteriophage. This results in display of the fused protein on the exterior of the phage, while the DNA encoding the fusion resides within the virion. The physical linkage between the displayed protein and the DNA encoding it allows screening of vast numbers of proteins for ligand-binding properties. With this technology, a phage clone encoding thioredoxin TrxA was isolated from a sub-library consisting of 80 clones. It is evident that the S. oneidensis clone set can be used for expression of functional S. oneidensis proteins in E. coli using the appropriate destination vectors. Characterization of ArcA. In Escherichia coli, metabolic transitions between aerobic and anaerobic growth states occur when cells enter an oxygen-limited condition. Many of these metabolic transitions are controlled at the transcriptional level by the activities of the global regulatory proteins ArcA (aerobic respiration control) and Fnr (fumarate nitrate regulator). A homolog of ArcA (81% amino acid sequence identity) was identified in S. oneidensis MR-1, and arcA mutants with MR-1 as the parental strain were generated. Phenotype characterization showed the arcA deletion mutant grew slower than the wild-type and was hypersensitive to H2O2 stress.

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号