A genome annotation-driven approach to cloning the human ORFeome

Genome Biol. 2004;5(10):R84. doi: 10.1186/gb-2004-5-10-r84. Epub 2004 Sep 30.

Abstract

We have developed a systematic approach to generating cDNA clones containing full-length open reading frames (ORFs), exploiting knowledge of gene structure from genomic sequence. Each ORF was amplified by PCR from a pool of primary cDNAs, cloned and confirmed by sequencing. We obtained clones representing 70% of genes on human chromosome 22, whereas searching available cDNA clone collections found at best 48% from a single collection and 60% for all collections combined.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromosomes, Human, Pair 22 / genetics
  • Cloning, Molecular / methods*
  • Computational Biology
  • DNA, Complementary / genetics
  • Databases, Genetic
  • Genome, Human*
  • Genomics / methods*
  • Humans
  • Open Reading Frames / genetics*
  • Polymerase Chain Reaction
  • Polymorphism, Single Nucleotide / genetics
  • Proteome / genetics*
  • Research Design
  • Sequence Analysis, DNA

Substances

  • DNA, Complementary
  • Proteome