SYNTHESIS/ASSEMBLY-BRANCHED CHAIN KETOACID DEHYDROGENASE
SYNTHESIS/ASSEMBLY-BRANCHED CHAIN KETOACID DEHYDROGENASE
批准号:
3237629
负责人:
DEAN J DANNER
金额:
$13.69万
依托单位:
依托单位国家:
美国
项目类别:
财政年份:
1986
资助国家:
美国
项目状态:
已结题
起止时间:
1986-08-15 至 1989-07-31
关键词:
complementary DNA enzyme complex fibroblasts gel electrophoresis gene expression gene mutation genetic library genetic manipulation human subject human tissue immunochemistry immunoelectrophoresis ketoacid maple syrup urine disease messenger RNA mitochondria monoclonal antibody oxidoreductase tissue /cell culture
中文摘要
总体目标是确定指导合成的基因
英文摘要
The overall goal is to define the genes directing the synthesis of the
mitochondrial enzyme complex, branched chain Alpha-ketoacid dehydrogenase
[BCKD]. Inherited human mutations are known which specifically affect the
function of BCKD known as Maple Syrup Urine Disease [MSUD]. These studies
will increase our understanding of the defect and provide improved
treatment for affected individuals. Immediate goals include: 1)
Immunochemical characterization of BCKD proteins in human cells expressing
defective function of the catalytic activity, 2) Biochemical
characterization of the cytosolic synthesis and mitochondrial processing of
the precursor proteins to BCKD, and 3) Construction of cDNA probes to study
the genes and mRNAs directing the synthesis of these proteins. Polyclonal
and monoclonal antibodies which specifically recognize the proteins of BCKD
are used to screen mitochondrial proteins from fibroblasts derived from
MSUD patients for cross-reactive material [CRM]. CRM- cells will be used
to quantify BCKD proteins by crossed-immunoelectrophoretic methods.
Western Blot analysis of two-dimensional gel separation of the
mitochondrial proteins will also be used to study the mutant proteins. In
vitro translation of RNA isolated from different sources, especially cells
induced to overproduce mitochondrial proteins, will be used for the
synthesis of these proteins. Processing studies will analysize conditions
required for uptake of in vitro translated proteins by freshly isolated,
respiratory competent mitochondria. Construction of cDNA probes involve
using Lambdagt11 expression vectors with a human liver cDNA library to
select clones producing the BCKD proteins. This involves to use of
monospecific antibodies in selection of the clones and expansion of the
selected clones in plasmid vectors.
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会议论文
MAPLE SYRUP URINE DISEASE
-
批准号:6565741
-
项目类别:
-
资助金额:$29.31万
-
财政年份:2001
-
负责人:DEAN J DANNER
-
依托单位:
MAPLE SYRUP URINE DISEASE
-
批准号:6586036
-
项目类别:
-
资助金额:$29.31万
-
财政年份:2001
-
负责人:DEAN J DANNER
-
依托单位:
MAPLE SYRUP URINE DISEASE
-
批准号:6415359
-
项目类别:
-
资助金额:$29.31万
-
财政年份:2000
-
负责人:DEAN J DANNER
-
依托单位:
MAPLE SYRUP URINE DISEASE
-
批准号:6113172
-
项目类别:
-
资助金额:$3.88万
-
财政年份:1998
-
负责人:DEAN J DANNER
-
依托单位:
MAPLE SYRUP URINE DISEASE
-
批准号:6244358
-
项目类别:
-
资助金额:$3.62万
-
财政年份:1997
-
负责人:DEAN J DANNER
-
依托单位:
MAPLE SYRUP URINE DISEASE
-
批准号:6274406
-
项目类别:
-
资助金额:$3.57万
-
财政年份:1997
-
负责人:DEAN J DANNER
-
依托单位:
SYNTHESIS/ASSEMBLY BRANCHED CHAIN KETOACID DEHYDROGENASE
-
批准号:2140465
-
项目类别:
-
资助金额:$21.73万
-
财政年份:1986
-
负责人:DEAN J DANNER
-
依托单位:
SYNTHESIS/ASSEMBLY BRANCHED CHAIN KETOACID DEHYDROGENASE
-
批准号:2140466
-
项目类别:
-
资助金额:$22.29万
-
财政年份:1986
-
负责人:DEAN J DANNER
-
依托单位:
SYNTHESIS/ASSEMBLY-BRANCHED CHAIN KETOACID DEHYDROGENASE
-
批准号:3237631
-
项目类别:
-
资助金额:$15.19万
-
财政年份:1986
-
负责人:DEAN J DANNER
-
依托单位:
SYNTHESIS/ASSEMBLY BRANCHED CHAIN KETOACID DEHYDROGENASE
-
批准号:2140467
-
项目类别:
-
资助金额:$22.81万
-
财政年份:1986
-
负责人:DEAN J DANNER
-
依托单位:
SYNTHESIS/ASSEMBLY BRANCHED CHAIN KETOACID DEHYDROGENASE
-
批准号:2458751
-
项目类别:
-
资助金额:$23.72万
-
财政年份:1986
-
负责人:DEAN J DANNER
-
依托单位:
SYNTHESIS/ASSEMBLY-BRANCHED CHAIN KETOACID DEHYDROGENASE
-
批准号:3237634
-
项目类别:
-
资助金额:$18.28万
-
财政年份:1986
-
负责人:DEAN J DANNER
-
依托单位:
SYNTHESIS/ASSEMBLY-BRANCHED CHAIN KETOACID DEHYDROGENASE
-
批准号:3237635
-
项目类别:
-
资助金额:$19.58万
-
财政年份:1986
-
负责人:DEAN J DANNER
-
依托单位:
SYNTHESIS/ASSEMBLY BRANCHED CHAIN KETOACID DEHYDROGENASE
-
批准号:2879365
-
项目类别:
-
资助金额:$8.91万
-
财政年份:1986
-
负责人:DEAN J DANNER
-
依托单位:
SYNTHESIS/ASSEMBLY-BRANCHED CHAIN KETOACID DEHYDROGENASE
-
批准号:3237630
-
项目类别:
-
资助金额:$17.8万
-
财政年份:1986
-
负责人:DEAN J DANNER
-
依托单位:
SYNTHESIS/ASSEMBLY-BRANCHED CHAIN KETOACID DEHYDROGENASE
-
批准号:3237636
-
项目类别:
-
资助金额:$20.42万
-
财政年份:1986
-
负责人:DEAN J DANNER
-
依托单位:
SYNTHESIS/ASSEMBLY-BRANCHED CHAIN KETOACID DEHYDROGENASE
-
批准号:3237633
-
项目类别:
-
资助金额:$18.28万
-
财政年份:1986
-
负责人:DEAN J DANNER
-
依托单位:
SYNTHESIS/ASSEMBLY-BRANCHED CHAIN KETOACID DEHYDROGENASE
-
批准号:3237632
-
项目类别:
-
资助金额:$14.97万
-
财政年份:1986
-
负责人:DEAN J DANNER
-
依托单位:
海外基金