Interrogating the transgenic genome: development of an interspecies tiling array

Syst Biol Reprod Med. 2011 Feb;57(1-2):54-62. doi: 10.3109/19396368.2010.506000. Epub 2011 Jan 10.

Abstract

A single expressing copy of the human protamine domain was randomly inserted into an intron of Cyp2c38. The transgenic locus was shown to recapitulate the level of expression observed in normal human testis while not perturbing endogenous protamine expression. The development of an interspecies tiling array was pursued to enable direct comparison of the orthologous protamine domains in a single experiment. Probe design was adapted to generate species-specific high resolution probe sets that would tolerate repetitive elements. Results from competitive hybridizations demonstrate that interspecies tiling arrays are a valuable tool for parallel analysis of highly similar DNA sequences. This approach provides a rapid and reliable means of interrogating samples prior to deep sequencing analysis. These arrays should readily compliment most DNA isolation and analysis techniques such as ChIP, nuclease sensitivity and nuclear matrix association assays.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cytochrome P-450 Enzyme System / genetics*
  • Cytochrome P450 Family 2
  • Humans
  • Male
  • Mice
  • Mice, Transgenic
  • Oligonucleotide Array Sequence Analysis / methods*
  • Protamines / genetics*
  • Testis / metabolism
  • Transgenes / genetics*

Substances

  • Protamines
  • Cytochrome P-450 Enzyme System
  • CYP2C38 protein, mouse
  • Cytochrome P450 Family 2