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Related Concept Videos

Reporter Genes02:11

Reporter Genes

Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
Commonly used reporter...
Cis-regulatory Sequences02:02

Cis-regulatory Sequences

Cis-regulatory sequences are short fragments of non-coding DNA that are present on the same chromosomes as the genes that they regulate. These fragments serve as binding sites for transcriptional regulators, proteins that are responsible for controlling gene transcription and differential gene expression across cell types in eukaryotes. Cis-regulatory sequences can be close to the gene of interest or thousands of bases away in the DNA sequence; however, those sequences that are further away are...
Cis-regulatory Sequences02:02

Cis-regulatory Sequences

Cis-regulatory sequences are short fragments of non-coding DNA that are present on the same chromosomes as the genes that they regulate. These fragments serve as binding sites for transcriptional regulators, proteins that are responsible for controlling gene transcription and differential gene expression across cell types in eukaryotes. Cis-regulatory sequences can be close to the gene of interest or thousands of bases away in the DNA sequence; however, those sequences that are further away are...
Cooperative Binding of Transcription Regulators02:13

Cooperative Binding of Transcription Regulators

Transcriptional regulators bind to specific cis-regulatory sequences in the DNA to regulate gene transcription. These cis-regulatory sequences are very short, usually less than ten nucleotide pairs in length. The short length means that there is a high probability of the exact same sequence randomly occurring throughout the genome.  Since regulators can also bind to groups of similar sequences, this further increases the chances of random binding. Transcriptional regulators form dimers that...

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Quantitative Comparison of cis-Regulatory Element (CRE) Activities in Transgenic Drosophila melanogaster
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Making reporter gene constructs to analyze cis-regulatory elements.

José Bessa1, José Luis Gómez-Skarmeta

  • 1Centro Andaluz de Biología del Desarrollo (CABD), CSIC-Universidad Pablo de Olavide, Seville, Spain. jcribbes@upo.es

Methods in Molecular Biology (Clifton, N.J.)
|November 9, 2011
PubMed
Summary

This study presents a new protocol for isolating cis-regulatory sequences from genomic DNA. This method facilitates functional analysis of gene transcription control elements.

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Area of Science:

  • Genetics
  • Molecular Biology
  • Developmental Biology

Background:

  • Cis-regulatory sequences are crucial for controlling gene transcription timing, location, and levels.
  • Understanding these elements is fundamental to comprehending development, cell differentiation, and morphogenesis.
  • Existing methods for cis-acting sequence detection include in silico analysis and ChIP-seq experiments.

Purpose of the Study:

  • To describe a novel protocol for the isolation of cis-regulatory sequences.
  • To enable functional assays of these regulatory elements.
  • To utilize Gateway cloning technology for efficient cloning into expression vectors.

Main Methods:

  • Isolation of cis-regulatory sequences from genomic DNA.
  • Cloning of isolated sequences into expression vectors.
  • Utilizing the Gateway cloning system for seamless vector integration.

Main Results:

  • A described protocol for isolating cis-regulatory sequences is provided.
  • The protocol facilitates the cloning of these sequences into expression vectors.
  • The method is amenable to functional assays for studying gene regulation.

Conclusions:

  • The presented protocol offers a new approach for studying cis-regulatory sequences.
  • This method aids in understanding gene transcription control.
  • The protocol supports functional characterization of regulatory elements through expression vector cloning.