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GENETIC ANALYSIS OF BACTERIOPHAGE T4 REGA PROTEIN-RNA

噬菌体 T4 REGA 蛋白-RNA 的遗传分析

基本信息

批准号：
3466370
负责人：
ERIC S MILLER
金额：
$ 0.43万
依托单位：
NORTH CAROLINA STATE UNIVERSITY RALEIGH
依托单位国家：
美国
项目类别：
财政年份：
1987
资助国家：
美国
起止时间：
1987-08-01 至 1994-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3466370
关键词：
Escherichia coli RNA bacteriophage T4 binding proteins gel electrophoresis gene expression gene induction /repression genetic translation messenger RNA mutant nucleic acid sequence plasmids protein sequence virus protein

项目摘要

The bacteriophage T4 regA protein is a translational repressor of several T4 mRNAs. Genetic methods are described in this proposal that will identify amino acids of the regA protein that contact the mRNA binding sites. In the principle project, plasmids that allow controlled regA expression will be mutagenized in vitro and in vivo and will then be screened for altered regA function in E. coli strains containing regA-sensitive genes or sensitive gene fusions. Kinetic descriptions will be obtained for regA proteins with diminished, tight-binding or new site-specifity phenotypes that will then be correlated to the single amino acid substitution (determined by nucleic acid sequencing) in each repressor mutant. Amino acid residues and short regions of regA have been selected for targeted mutagenesis. From the initial results using random mutagenesis and the described new selection techniques, regA residues of particular interest will be identified for further directed mutagenesis studies. Two aspects of regA function are addressed as a smaller effort of this proposal. A translational activation function has been tentatively ascribed to the regA protein; the activated proteins will be identified by 2D gel electrophoresis and sequenced. The regA mRNA binding site will also be characterized by random oligonucleotide synthesis in a collaborative effort with an established laboratory working on translational regulation. These efforts will complement our principle interests described above. The long-term goal of the laboratory is to define the RNA-protein interactions of the regA translational repressor with the expectation that features of regA will be relevant to understanding RNA-protein interactions of other prokaryotic and eukaryotic systems.

噬菌体T4 regA蛋白是一种翻译抑制因子

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

RNA-protein interactions of the bacteriophage RB69 RegA translational repressor protein.

噬菌体 RB69 RegA 翻译抑制蛋白的 RNA-蛋白相互作用。

DOI：
发表时间：
1995
期刊：
Nucleic acids symposium series.
影响因子：
0
作者：
Jozwik,CE;Miller,ES
通讯作者：
Miller,ES

Sequence analysis of conserved regA and variable orf43.1 genes in T4-like bacteriophages.

T4 样噬菌体中保守 regA 和可变 orf43.1 基因的序列分析。

DOI：
10.1128/jb.172.9.5180-5186.1990
发表时间：
1990
期刊：
Journal of bacteriology
影响因子：
3.2
作者：
Miller,ES;Jozwik,CE
通讯作者：
Jozwik,CE

Regions of bacteriophage T4 and RB69 RegA translational repressor proteins that determine RNA-binding specificity.

决定 RNA 结合特异性的噬菌体 T4 和 RB69 RegA 翻译抑制蛋白区域。

DOI：
10.1073/pnas.89.11.5053
发表时间：
1992
期刊：
Proceedings of the National Academy of Sciences of the United States of America
影响因子：
11.1
作者：
Jozwik,CE;Miller,ES
通讯作者：
Miller,ES

An E. coli B mutation, rpoB5081, that prevents growth of phage T4 strains defective in host DNA degradation.

大肠杆菌 B 突变 rpoB5081 可阻止宿主 DNA 降解缺陷的噬菌体 T4 菌株的生长。