用一种高效复制和转录的非自然基对类型对DNA和RNA进行特定地点的标记
Young Jun Seo1, Denis A Malyshev, Thomas Lavergne
1Department of Chemistry and Center for Protein and Nucleic Acid Research, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, California 92037, USA.
Journal of the American Chemical Society
|October 11, 2011
概括
研究人员开发了新的非自然基对,用于特定站点的DNA和RNA标签. 这些改性核酸使得高效的酶合成和合成后功能化为各种应用.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 合成生物学 合成生物学
背景情况:
- 核酸的特定地点标签对于生物物理学,核酸进化,纳米材料和生物传感器的应用至关重要.
- 将遗传字母扩展到不自然的基因对,有助于创建新的标签策略.
研究的目的:
- 合成和分析经过修改的非自然基对, (d) 5SICS和 (d) MMO2,用propargylamine链接剂进行特定部位的DNA和RNA修改.
- 为了评估一个α-酸酸盐变体的脊柱化和后放大标签.
- 评估这些修饰核酸在酶合成中的效率和忠实性.
主要方法:
- 合成具有功能链接器的d5SICS和dMMO2的ribos和deoxyribo变体.
- 使用稳定状态动力学和聚合酶链反应 (PCR) 进行脱氧核酸的表征.
- 通过模型RNA和tRNA的转录对核糖核酸的表征.
主要成果:
- 成功合成和表征修饰的 (d) 5SICS和 (d) MMO2核酸.
- 在酶合成期间或之后证明了DNA和RNA的高效和特定位置的标记.
- 鉴定了核酸和聚合酶对链接器附着的特定敏感性,但总体上实现了高效率和保真性.
结论:
- 开发出的非自然基对 (d) 5SICS和 (d) MMO2 适合生产特定位点的修饰DNA和RNA.
- 这些经过修改的核酸可以与多个多样化的组合进行功能化,以实现实际应用.
- 这些策略为高级研究和生物技术的核酸标签提供了多功能途径.
相关概念视频
Labeling DNA Probes
DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
In-situ Hybridization
In situ hybridization (ISH) is a technique used to detect and localize specific DNA or RNA molecules in cells, tissue, or tissue sections using a labeled probe. The technique was first used in 1969 for the investigation of nucleic acids. It is currently an essential tool in scientific research and clinical settings, especially for diagnostic purposes.
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...
Conservative Site-specific Recombination and Phase Variation
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...
Sanger Sequencing
DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...
Maxam-Gilbert Sequencing
In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
Challenges of the Maxam-Gilbert Method
The...
Challenges of the Maxam-Gilbert Method
The...
Next-generation Sequencing
The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features.


