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Related Experiment Videos

Mining the human genome using microarrays of open reading frames.

S G Penn1, D R Rank, D K Hanzel

  • 1Advanced Research Team, Molecular Dynamics Inc., Sunnyvale, California, USA.

Nature Genetics
|November 4, 2000
PubMed
Summary

The Human Genome Project reveals many novel genes missed by expressed sequence tags (ESTs). Genomic sequencing is crucial for discovering the full spectrum of human genes.

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Area of Science:

  • Genomics
  • Molecular Biology
  • Bioinformatics

Background:

  • Expressed sequence tags (ESTs) have been instrumental in gene discovery.
  • The Human Genome Project aims to comprehensively map human genes.
  • The extent of gene discovery via ESTs versus genomic sequencing remains an open question.

Purpose of the Study:

  • To test if the Human Genome Project uncovers genes missed by EST sequencing.
  • To identify novel human genes directly from genomic sequence using gene expression data.

Main Methods:

  • Designed and produced microarrays with probes from open reading frames (ORFs) in human genomic sequence.
  • Analyzed genomic sequence using ORF prediction programs.
  • Amplified ORFs from genomic DNA and arrayed them for gene expression analysis.

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Main Results:

  • Of the first 10,000 arrayed ORFs, 31% were novel and 29% were similar but not identical to known sequences.
  • Approximately 50% of these novel/similar ORFs were expressed in queried tissues.
  • Subsequent verification confirmed expression of several novel genes.

Conclusions:

  • Genomic sequencing is essential for discovering many human genes not identified through ESTs.
  • ESTs alone are insufficient for a complete catalog of human genes.
  • The Human Genome Project is vital for comprehensive human gene discovery.