DNA-BINDING PROTEINS IN TRYPANOSOMA BRUCEI

布氏锥虫中的 DNA 结合蛋白

• 批准号: 3137470
• 负责人: DAVID A CAMPBELL
• 金额: $ 11.41万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-07-01 至 1992-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3137470
• 关键词: DNA binding protein; DNA footprinting; Trypanosoma; autoradiography; developmental genetics; endonuclease; gel electrophoresis; gene expression; genes; genetic mapping; genetic promoter element; genetic transcription; laboratory mouse; laboratory rat; molecular cloning; nuclear runoff assay; nucleic acid sequence; nucleic acid structure; protein structure

Potential targets for chemotherapy in microbial infections are the biochemical pathways which are unique to the pathogen. For the African trypanosomes, the obligatory synthesis of all messenger RNAs by dicontinuous transcription represents a target for such a strategy. The long term aim of this research is to analyze gene structure and discontinuous transcription in Trypanosoma brucei. The experiments described here will define protein-DNA interactions in the proximity of the start sites of transcription of a) the mini- exon) (5'-exon) and b) representative protein-coding genes (3'- exons). These two-classes of genes are representative of the two discrete genetic loci involved in discontinuous transcription. Gel- retardation and Exonuclease III-protection assays will be used to identify and characterize specific protein-DNA interactions on a cloned mini-exon gene incubated with crude nuclear extracts from T. brucei. Nuclear extracts enriched by chromatography for the specific DNA-binding proteins will be used in DNA footprinting assays to identify the nucleotides bound by the protein. The transcription initiation sites of active ubiquitin genes and the 117 VSG gene, two apparently different classes of 3'-exon, will be identified by cosmid cloning, the analysis of nascent RNA by nuclear run-on and nuclease-protection assays. Gel retardation assays will be used to characterize proteins binding to the transcription indication sites of these 3'-exons.

微生物感染化疗的潜在靶点是

项目成果

c2X75, a derivative of the cosmid vector c2XB.

c2X75，粘粒载体 c2XB 的衍生物。

• DOI: 10.1093/nar/17.1.458
• 发表时间: 1989
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Campbell,DA

Ubiquitin-EP52 fusion protein homologs from Trypanosoma brucei.

来自布氏锥虫的泛素-EP52 融合蛋白同源物。

• DOI: 10.1093/nar/18.23.7181
• 发表时间: 1990
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Wong,S;Morales,TH;Campbell,DA
• 通讯作者: Campbell,DA

A polyubiquitin gene from Trypanosoma brucei.

来自布氏锥虫的多聚泛素基因。

• DOI: 10.1016/0166-6851(89)90111-4
• 发表时间: 1989
• 期刊: Molecular and biochemical parasitology
• 影响因子: 1.5
• 作者: Wong,S;Campbell,DA
• 通讯作者: Campbell,DA