Isolation of human transcribed sequences from human-rodent somatic cell hybrids

Science. 1989 Nov 10;246(4931):813-5. doi: 10.1126/science.2479099.

Abstract

A method was developed for selectively isolating genes from localized regions of the human genome that are contained in interspecific hybrid cells. Complementary human DNA was prepared from a human-rodent somatic cell hybrid that contained less than 1% human DNA, by using consensus 5' intron splice sequences as primers. These primers would select immature, unspliced messenger RNA (still retaining species-specific repeat sequences) as templates. Screening a derived complementary DNA library for human repeat sequences resulted in the isolation of human clones at the anticipated frequency with characteristics expected of exons of transcribed human genes--single copy sequences that hybridized to discrete bands on Northern (RNA) blots.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Blotting, Northern
  • Blotting, Southern
  • Chromosome Mapping
  • Chromosomes, Human, Pair 19
  • Cloning, Molecular
  • Cricetinae
  • DNA / biosynthesis
  • DNA / genetics
  • DNA / isolation & purification*
  • Humans
  • Hybrid Cells*
  • Introns
  • Nucleic Acid Hybridization
  • RNA / genetics
  • Repetitive Sequences, Nucleic Acid
  • Restriction Mapping
  • Templates, Genetic

Substances

  • RNA
  • DNA