PHAGE T4 HEAD ASSEMBLY AND INITIATION OF INFECTION
PHAGE T4 HEAD ASSEMBLY AND INITIATION OF INFECTION
批准号:
2059805
负责人:
LINDSAY W BLACK
金额:
$27.03万
依托单位国家:
美国
项目类别:
财政年份:
1977
资助国家:
美国
项目状态:
已结题
起止时间:
1977-07-01 至 1997-02-28
关键词:
DNA virus affinity labeling bacterial genetics bacteriophage T4 capsid conformation crosslink enzyme mechanism fusion gene host organism interaction membrane proteins microorganism growth microorganism metabolism molecular pathology mutant protein biosynthesis protein sequence virus DNA virus envelope virus genetics virus infection mechanism
中文摘要
研究重点为病毒DNA包装机制
以及噬菌体T4体外包装衍生的构建
克隆载体。 我们的工作特别针对 1)
了解多功能的结构和功能
噬菌体 T4 包装(前头和终止酶)蛋白。 3-
酶促(DNA 易位)的维度基础和
主要衣壳的结构(前头形状决定)作用
蛋白质 gp23*),是一种微加工酶产物 (gp23**),
以及衣壳基因突变体的数量将被确定。 的组装
DNA 入口(也是前头起始)顶点作为
将使用过表达来检查整合膜蛋白
载体;催化基因产物和宿主系统的作用
将确定膜插入。 具体包装
这两个前头蛋白的突变都将被选择
基因定向诱变后。 DNA 终止酶
与 DNA 入口顶点蛋白相互作用的蛋白质
前头已被过表达和纯化-它们
DNA 易位和串联体的作用机制
切割将被调查。 lambda 和 lambda-pBR322
衍生的 DNA 可以在体外包装到 T4 头中。 因斯
和/或 P1 DNA 包装到 T4 中并用
同源位点特异性重组系统应允许
T4-杂交大粘粒载体的开发。 我们提出了
一种新颖的“螺旋折叠”包装噬菌体 DNA 模型。
形态学和(噬菌体)化学工作可以明确地
建立这个模型。 包装的扭结或非 B 型 DNA 将
检测到,及其与 DNA 结合位点的相互作用
主要衣壳蛋白及其次要加工酶产物
(gp23**) 将被探测。 我们将确定 DNA 结构
可体内、体外包装;例如 是否有刻痕和
异源双链体环从体外包装的 DNA 中排除,并且
终止酶蛋白是否起到区分这种作用
耦合 DNA 修复过程中的结构。
英文摘要
The research focuses on the mechanism of viral DNA packaging
and on the construction of phage T4 in vitro packaging-derived
cloning vectors. Our work is especially directed toward 1)
understanding the structures and functions of the multifunctional
phage T4 packaging (prohead and terminase) proteins. The 3-
dimensional basis for enzymatic (DNA translocating) and
structural (prohead form-determining) roles of the major capsid
protein gp23*), of a minor processed enzymatic product (gp23**),
and of capsid gene mutants will be determined. The assembly of
the DNA entrance (also the prohead initiation) vertex as an
integral membrane protein will be examined using overexpression
vectors; the role of a catalytic gene product and host systems in
the membrane insertion will be determined. Specific packaging
mutations in both these prohead proteins will be selected
following gene-directed mutagenesis. The DNA terminase
proteins which interact with the DNA entrance vertex protein in
the prohead have been overexpressed and purified-their
mechanisms of action in DNA translocation and concatemer
cutting will be investigated. lambda and lambda-pBR322
derivative DNAs can be packaged into T4 heads in vitro. cos
and/or P1 DNAs packaged into T4 and recircularized with the
homologous site-specific recombination systems should allow
development of T4-hybrid megacosmid vectors. We have proposed
a novel "spiral-fold" model for packaged phage DNA.
Morphological and (with phage ) chemical work can definitively
establish this model. Packaged kinked or non-B form DNA will be
detected, and its interaction with the DNA binding site in the
major capsid protein and its minor processed enzymatic product
(gp23**) will be probed. We will determine what DNA structures
can be packaged in vivo and in vitro; e.g. whether nicks and
heteroduplex loops are excluded from packaged DNA in vitro, and
whether the terminase proteins act to discriminate against such
structures in coupled DNA repair processes.
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会议论文
Mechanism of bacteriophage DNA packaging initiation and DNA translocation.
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批准号:9274832
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项目类别:
-
资助金额:$30.51万
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财政年份:2016
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负责人:LINDSAY W BLACK
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依托单位:
Mechanism of bacteriophage DNA packaging initiation and DNA translocation.
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批准号:9080621
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项目类别:
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资助金额:$30.42万
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财政年份:2016
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负责人:LINDSAY W BLACK
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依托单位:
PHAGE T4 HEAD ASSEMBLY AND INITIATION OF INFECTION
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批准号:2059806
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项目类别:
-
资助金额:$28.11万
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财政年份:1977
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负责人:LINDSAY W BLACK
-
依托单位:
PHAGE T4 HEAD ASSEMBLY AND INITIATION OF INFECTION
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批准号:3480617
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项目类别:
-
资助金额:$24.24万
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财政年份:1977
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负责人:LINDSAY W BLACK
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依托单位:
Phage T4 Head Assembly and Initiation of Infection
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批准号:8427269
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项目类别:
-
资助金额:$34.55万
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财政年份:1977
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负责人:LINDSAY W BLACK
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依托单位:
Phase T4 Head Assembly and Initiation of Infection
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批准号:6858575
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项目类别:
-
资助金额:$37.13万
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财政年份:1977
-
负责人:LINDSAY W BLACK
-
依托单位:
PHAGE T4 HEAD ASSEMBLY AND INITIATION OF INFECTION
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批准号:3480623
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项目类别:
-
资助金额:$26.81万
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财政年份:1977
-
负责人:LINDSAY W BLACK
-
依托单位:
PHAGE T4 HEAD ASSEMBLY AND INITIATION OF INFECTION
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批准号:3480620
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项目类别:
-
资助金额:$19.63万
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财政年份:1977
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负责人:LINDSAY W BLACK
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依托单位:
Phase T4 Head Assembly and Initiation of Infection
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批准号:6780511
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项目类别:
-
资助金额:$18.56万
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财政年份:1977
-
负责人:LINDSAY W BLACK
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依托单位:
PHAGE T4 HEAD ASSEMBLY AND INITIATION OF INFECTION
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批准号:3480622
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项目类别:
-
资助金额:$21.17万
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财政年份:1977
-
负责人:LINDSAY W BLACK
-
依托单位:
PHAGE T4 HEAD ASSEMBLY AND INITIATION OF INFECTION
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批准号:2671667
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项目类别:
-
资助金额:$30.96万
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财政年份:1977
-
负责人:LINDSAY W BLACK
-
依托单位:
PHAGE T4 HEAD ASSEMBLY AND INITIATION OF INFECTION
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批准号:6372951
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项目类别:
-
资助金额:$33.69万
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财政年份:1977
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负责人:LINDSAY W BLACK
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依托单位:
Phage T4 Head Assembly and Initiation of Infection
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批准号:6681705
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项目类别:
-
资助金额:$37.13万
-
财政年份:1977
-
负责人:LINDSAY W BLACK
-
依托单位:
Phage T4 Head Assembly and Initiation of Infection
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批准号:7760640
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项目类别:
-
资助金额:$37.13万
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财政年份:1977
-
负责人:LINDSAY W BLACK
-
依托单位:
Phage T4 Head Assembly and Initiation of Infection
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批准号:8220775
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项目类别:
-
资助金额:$36.75万
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财政年份:1977
-
负责人:LINDSAY W BLACK
-
依托单位:
PHAGE T4 HEAD ASSEMBLY AND INITIATION OF INFECTION
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批准号:2886339
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项目类别:
-
资助金额:$31.79万
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财政年份:1977
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负责人:LINDSAY W BLACK
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依托单位:
PHAGE T4 HEAD ASSEMBLY AND INITIATION OF INFECTION
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批准号:2003152
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项目类别:
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资助金额:$30.06万
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财政年份:1977
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负责人:LINDSAY W BLACK
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依托单位:
PHAGE T4 HEAD ASSEMBLY AND INITIATION OF INFECTION
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批准号:6169355
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项目类别:
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资助金额:$32.71万
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财政年份:1977
-
负责人:LINDSAY W BLACK
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依托单位:
PHAGE T4 HEAD ASSEMBLY AND INITIATION OF INFECTION
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批准号:2059804
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项目类别:
-
资助金额:$26.05万
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财政年份:1977
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负责人:LINDSAY W BLACK
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依托单位:
PHAGE T4 HEAD ASSEMBLY AND INITIATION OF INFECTION
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批准号:3480621
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项目类别:
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资助金额:$20.35万
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财政年份:1977
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负责人:LINDSAY W BLACK
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依托单位:
海外基金