Characterization of expressed sequence tags generated from skin cDNA clones of Equus caballus by single pass sequencing

Anim Biotechnol. 2001 May;12(1):87-97. doi: 10.1081/ABIO-100102981.

Abstract

A cDNA library was built using RNA extracted from the skin tissue of an adult horse. The library was primed with oligo (dT) and sequences were directionally inserted in order to produce an expression library. The library has 5.8X 10(5) plaque forming units with 99.6% recombinant phage. The average insert size is 1.3 Kbp. Three hundred and thirteen expressed sequence tags (ESTs) were generated from sequencing of the 5 prime end of randomly selected skin cDNA clones. The ESTs were sequenced on an ABI 377 using Big-Dye chemistry. A similarity search was performed on each EST using the NCBI non-redundant protein database and 206 ESTs were putatively identified. Twenty six percent of the identified ESTs were redundant. The ESTs were categorized by function. The most frequently identified functional class was translational proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cloning, Molecular
  • DNA, Complementary / genetics*
  • Databases, Factual
  • Expressed Sequence Tags*
  • Gene Expression Regulation
  • Gene Library*
  • Horses / genetics*
  • Humans
  • Polymerase Chain Reaction
  • Protein Biosynthesis
  • Sequence Alignment
  • Sequence Analysis, DNA
  • Signal Transduction
  • Skin

Substances

  • DNA, Complementary