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Related Experiment Videos

Functional cloning, sorting, and expression profiling of nucleic acid-binding proteins.

Y Ramanathan1, Haibo Zhang, Virginie Aris

  • 1Center for Applied Genomics, Public Health Research Institute, International Center for Public Health W420M, Newark, New Jersey 07103, USA.

Genome Research
|August 15, 2002
PubMed
Summary

Researchers identified 170 human nucleic acid-binding proteins (NBPs) using a functional expression-cloning strategy. This study advances understanding of gene regulation and genome integrity by characterizing novel NBPs and their expression patterns.

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Area of Science:

  • Genomics
  • Molecular Biology
  • Bioinformatics

Background:

  • Elucidating gene function is crucial post-genome sequencing.
  • Proteins regulating gene expression or genome structure are key targets.
  • Identifying novel nucleic acid-binding proteins (NBPs) is essential for understanding cellular processes.

Purpose of the Study:

  • To develop and validate a functional expression-cloning strategy for identifying human NBPs.
  • To characterize the spectrum of NBPs and their binding properties.
  • To explore gene expression patterns and regulatory mechanisms of NBPs.

Main Methods:

  • Modified functional expression-cloning using oligonucleotide-binding substrates.
  • Screening of phage libraries from human colorectal adenocarcinoma and fetal lung tissues.

Related Experiment Videos

  • Complementary sorting strategies for NBP classification without protein purification.
  • GeneChip expression profiling across 14 breast-derived cell lines.
  • Cluster analysis to identify co-expressed genes and upstream regulatory region analysis.

    • Main Results:

    • Identification of approximately 170 human cDNAs encoding NBPs.
    • Sequence analysis confirmed known DNA- or RNA-binding motifs in identified clones.
    • A sorting strategy differentiated NBP classes based on binding properties.
    • Expression profiling revealed transcriptional activity patterns for identified NBPs and related genes.
    • Strategies for analyzing transcription factor binding sites in synexpression groups were presented.

    Conclusions:

    • The functional expression-cloning strategy is effective for identifying human NBPs.
    • The study provides a valuable resource of NBPs and insights into their functional roles.
    • Gene expression profiling and cluster analysis aid in understanding NBP regulation and function.