相关实验视频

Updated: Aug 1, 2026

Avidity-based Extracellular Interaction Screening (AVEXIS) for the Scalable Detection of Low-affinity Extracellular Receptor-Ligand Interactions

Avidity-based Extracellular Interaction Screening (AVEXIS) for the Scalable Detection of Low-affinity Extracellular Receptor-Ligand Interactions

Published on: March 5, 2012

增强剂结合蛋白的分子克隆:通过对表达库的选进行隔离,具有识别站点DNA的DNA

H Singh¹, J H LeBowitz, A S Baldwin

¹Center for Cancer Research, Massachusetts Institute of Technology, Cambridge 02139.

|February 12, 1988

概括

研究人员发现了一种新的DNA结合域,其特性类似于哺乳动物转录因子H2TF1和NF-kappa B. 这一发现有助于理解基因调节和蛋白质分析.

科学领域:

分子生物学分子生物学
遗传学是一个遗传学.
生物化学生物化学

背景情况:

像H2TF1和NF-kappa B这样的哺乳动物转录因子通过DNA结合来调节基因表达.
这些因素在基因增强剂中识别特定的DNA序列,例如主要基因相容性复合体 (MHC) I类和免疫球蛋白卡帕链基因中的特定DNA序列.
细胞类型分布和结合亲缘关系的差异区分这些转录因子.

研究的目的:

开发一种新的策略来分离编码DNA结合域的cDNA克隆.
描述一种新发现的DNA结合域,其与已知的转录因子重叠的识别特性.
为了研究已识别的DNA结合域的基因表达模式.

主要方法:

使用MHC增强器的DNA结合位点探针选一个兰巴巴菌体表达库.
描述菌体编码的融合蛋白的结合特异性,以增强序列.
分析分离的cDNA克隆的基因拷贝数和mRNA表达.

主要成果:

一个编码DNA结合域的新型cDNA克隆被成功分离出来.
编码的蛋白质表现出对MHC和kappa基因增强剂的特定结合.
该cDNA对应于一个单拷贝基因,表达为B细胞和非B细胞中的10kbmRNA.
已识别的DNA结合域与H2TF1和NF-kappa B.共享识别属性.

结论:

这种新的策略对于克隆和分析特定序列的DNA结合蛋白质是有效的.
隔离的DNA结合域代表了基因表达的潜在重要的调节器.
了解这个域的功能可以提供对转录因子识别机制的见解.

更多相关视频

Cell Surface Receptor Identification Using Genome-Scale CRISPR/Cas9 Genetic Screens

Cell Surface Receptor Identification Using Genome-Scale CRISPR/Cas9 Genetic Screens

Published on: June 6, 2020

AAV Deployment of Enhancer-Based Expression Constructs In Vivo in Mouse Brain

AAV Deployment of Enhancer-Based Expression Constructs In Vivo in Mouse Brain

Published on: March 31, 2022

相关概念视频

Labeling DNA Probes

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...

Southern Blot

Southern Blot

Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...