DESIGN AND SYNTHESIS OF NUCLEOTIDE ANALOGUES
核苷酸类似物的设计与合成
基本信息
- 批准号:6269685
- 负责人:
- 金额:$ 21.36万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:1998
- 资助国家:美国
- 起止时间:1998-02-01 至 1999-01-31
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
(Applicant's Description) An important goal of the program project is the
development of a high sensitivity polymerase chain reaction/restriction
endonuclease/ligase detection reaction (PCR/RE/LDR) method to identify
specific mutations in the presence of excess normal DNA (Sensitivity of 1
in 100,000 - 1,000,000 cells). To this end, a new class of nucleotide
analogues herein termed "convertides", have been designed to provide a
means to convert a wild-type sequence into one which contains a
restriction endonuclease recognition site. Convertides are defined as
nucleoside analogues that have base modifications or replacements which
allow them to pair to one or more of the natural bases in hybridization
step ("read"), and also to function as a template for another base in a
DNA replication reaction ("write"). A successful convertide will pair to
one or more natural bases when annealing to a target, allowing for
efficient extension by Tag polymerase (read), and also functions as a
template for incorporation of another base the tag polymerase copies the
primer-containing strand (write).
For performing the 12 possible base conversions, 7 known
deoxyribonucleoside analogues (Q1, Q2, Q4-Q8) and 13 newly-designed,
modified deoxyribonucleosides (Q3, Q9-20) will be investigated. Specific
objectives include: Synthetic routes to deoxyribonucleosides Q3, Q9-20
and their characterization. ([1] H and [13] C NMR, mass spectrometry,
absorption spectroscopy, and elemental analysis; Synthesis of 5'-
Dimethoxytrityl (DMT)-protected derivatives of the convertides will be
prepared and transformed to 3'-phophoramidites and 3'-linked CPG solid
supports for incorporation into oligonucleotides for Projects 1 and 3. The
effect of the modified nucleotides on duplex structure will be determined
by measuring and analyzing the helix-coil transition of sets of duplexes
containing each of the Q nucleosides opposite each of the natural
nucleosides. Synthesis of primers with backbone (phosphate and sugar)
modifications at or near the 3'-end will be explored for: (1) preventing
cleavage of 3'-convertide by proofreading polymerases in PCR reactions
(Project 1) and (2) improving fidelity of ligase reactions by
destabilization of ligase-DNA complex (Project 3).
(申请人描述)本项目的一个重要目标是
项目成果
期刊论文数量(0)
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专利数量(0)
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Donald E. BERGSTROM其他文献
Donald E. BERGSTROM的其他文献
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{{ truncateString('Donald E. BERGSTROM', 18)}}的其他基金
MODIFIED NUCLEOSIDES AS TOOLS FOR MOLECULAR BIOLOGY
修饰核苷作为分子生物学的工具
- 批准号:
6180751 - 财政年份:1996
- 资助金额:
$ 21.36万 - 项目类别:
MODIFIED NUCLEOSIDES AS TOOLS FOR MOLECULAR BIOLOGY
修饰核苷作为分子生物学的工具
- 批准号:
2685067 - 财政年份:1996
- 资助金额:
$ 21.36万 - 项目类别:
MODIFIED NUCLEOSIDES AS TOOLS FOR MOLECULAR BIOLOGY
修饰核苷作为分子生物学的工具
- 批准号:
2192442 - 财政年份:1996
- 资助金额:
$ 21.36万 - 项目类别:
MODIFIED NUCLEOSIDES AS TOOLS FOR MOLECULAR BIOLOGY
修饰核苷作为分子生物学的工具
- 批准号:
2392246 - 财政年份:1996
- 资助金额:
$ 21.36万 - 项目类别:
MODIFIED NUCLEOSIDES AS TOOLS FOR MOLECULAR BIOLOGY
修饰核苷作为分子生物学的工具
- 批准号:
6386205 - 财政年份:1996
- 资助金额:
$ 21.36万 - 项目类别:
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