Cloning of TPA-inducible early (TIE) genes by differential hybridization using TPA-Nonresponsive variant of mouse 3T3-L1 cells

Masayuki Amagai, Yoshio Inokuchi, Takeji Nishikawa, Yoshiko Shimizu, Nobuyoshi Shimizu

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The tumor promoter 12-O -tetradecanoylphorbol-13-acetate (TPA) induces DNA synthesis in quiescent 3T3-L1 cells but not in its variant VT-1 cells. A λgt10 cDNA library was constructed using poly(A)+ RNA from 3T3-L1 cells that were stimulated by TPA for 20 min. Radioactive cDNA probes were prepared from mRNAs of TPA-treated 3 T3-L1 and VT-1 cells and used for screening of the 3T3-L1 cDNA library by differential hybridization. Nine of 6000 phage plaques hybridized only to the 3T3-L1 cDNA probe. Analysis of the nucleotide sequence of five of these clones indicated a high degree of homology with human or mouse type I and type III collagen genes. Three other independent clones showed no homology with any known DNA sequences. These isolated clones of TPA-inducible early (TIE) genes may be useful to study the signal transduction pathway of phorbol esters.

Original languageEnglish
Pages (from-to)153-158
Number of pages6
JournalSomatic Cell and Molecular Genetics
Volume15
Issue number2
DOIs
Publication statusPublished - 1989 Mar

Fingerprint

3T3-L1 Cells
Tetradecanoylphorbol Acetate
Organism Cloning
Clone Cells
Gene Library
Genes
Complementary DNA
Messenger RNA
Collagen Type III
Phorbol Esters
Collagen Type I
Carcinogens
Bacteriophages
Signal Transduction
DNA

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

Cloning of TPA-inducible early (TIE) genes by differential hybridization using TPA-Nonresponsive variant of mouse 3T3-L1 cells. / Amagai, Masayuki; Inokuchi, Yoshio; Nishikawa, Takeji; Shimizu, Yoshiko; Shimizu, Nobuyoshi.

In: Somatic Cell and Molecular Genetics, Vol. 15, No. 2, 03.1989, p. 153-158.

Research output: Contribution to journalArticle

Amagai, Masayuki ; Inokuchi, Yoshio ; Nishikawa, Takeji ; Shimizu, Yoshiko ; Shimizu, Nobuyoshi. / Cloning of TPA-inducible early (TIE) genes by differential hybridization using TPA-Nonresponsive variant of mouse 3T3-L1 cells. In: Somatic Cell and Molecular Genetics. 1989 ; Vol. 15, No. 2. pp. 153-158.
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