Isolation of growth suppressors from a cDNA expression library

Oncogene. 1998 Dec 17;17(24):3187-97. doi: 10.1038/sj.onc.1202260.

Abstract

We describe an experimental procedure for the isolation of growth inhibitory sequences from a complex cDNA library. This approach first takes advantage of the SETGAP technique (selectable expression of transient growth arrest phenotype) to enrich for growth inhibitory sequences, followed by a screening procedure to identify individual cDNAs that inhibit cell proliferation. Here we provide a detailed description of the experimental protocol and report the characterization of two cDNA sequences isolated in our initial screen of a mouse cDNA library. One of these cDNAs encodes the mouse ubiquitin-conjugation enzyme UbcM2. The other encodes a truncated form of a novel WD40 repeat protein, named Bopl, which is conserved from yeast to human. Together, these results demonstrate a new approach for the isolation of growth suppressors from cDNA libraries, and identify a previously unknown gene likely to be involved in growth control.

Publication types

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

MeSH terms

  • 3T3 Cells
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cells, Cultured
  • DNA, Complementary
  • Fungal Proteins / genetics*
  • G1 Phase
  • Gene Expression
  • Gene Library
  • Growth Inhibitors / genetics*
  • Humans
  • Ligases / genetics*
  • Mice
  • Molecular Sequence Data
  • Saccharomyces cerevisiae Proteins*
  • Sequence Homology, Amino Acid
  • Ubiquitin-Conjugating Enzymes

Substances

  • DNA, Complementary
  • FLC1 protein, S cerevisiae
  • Fungal Proteins
  • Growth Inhibitors
  • Saccharomyces cerevisiae Proteins
  • Ubiquitin-Conjugating Enzymes
  • Ligases

Associated data

  • GENBANK/AF003346
  • GENBANK/U77415