聚合酶连锁反应和转录使用锁定核酸核酸三酸盐
Rakesh N Veedu1, Birte Vester, Jesper Wengel
1Nucleic Acid Center, Department of Physics and Chemistry, University of Southern Denmark, Campusvej 55, Odense M, 5230, Denmark.
Journal of the American Chemical Society
|June 7, 2008
概括
锁定核酸 (LNA) 改性DNA链可以使用聚合酶链反应放大,并通过DNA和RNA聚合酶转录,从而使新的分子生物学应用成为可能.
科学领域:
- 分子生物学分子生物学
- 核酸化学的核酸化学
背景情况:
- 锁定核酸 (LNA) 是化学修饰的DNA类似物,具有增强的结合亲和力和稳定性.
- 将LNA集成到DNA和RNA合成中,对酶性操纵提出了挑战.
研究的目的:
- 研究LNA修饰DNA链的酶放大和转录的可行性.
- 评估LNA修饰基质与标准DNA和RNA聚合酶的兼容性.
主要方法:
- 聚合酶链反应 (PCR) 用于放大含有LNA修改的DNA链.
- 实验室转录反应使用LNA-A核酸5'-三酸与各种DNA和RNA聚合酶进行.
主要成果:
- 通过PCR实现了通过LNA修饰的DNA链的成功放大.
- 通过DNA和RNA聚合酶成功完成了利用LNA-A核酸5'-三酸的转录反应.
结论:
- 对LNA修饰核酸的酶性操纵是可行的.
- DNA和RNA聚合酶可以有效地处理含有LNA的基质,扩大它们在分子生物学中的实用性.
相关概念视频
PCR
Overview
Real Time RT-PCR
Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
The real-time quantification of the number of amplified products is...
Bacterial Transcription
RNA polymerase (RNAP) carries out DNA-dependent RNA synthesis in both bacteria and eukaryotes. Bacteria do not have a membrane-bound nucleus. So, transcription and translation occur simultaneously, on the same DNA template.
Transcription can be divided into three main stages, each involving distinct DNA sequences to guide the polymerase. These are:
Transcription can be divided into three main stages, each involving distinct DNA sequences to guide the polymerase. These are:
PCR - Polymerase Chain Reaction
Overview
Bacterial RNA Polymerase
Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
Translesion DNA Polymerases
Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...


