Molecular cloning and characterization of an activated human c-raf-1 gene

M. Fukui, T. Yamamoto, S. Kawai, Fumihiro Mitsunobu, K. Toyoshima

Research output: Contribution to journalArticle

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Abstract

Results of previous studies have shown that a raf-related transforming DNA sequence is present in NIH 3T3 transformants that are derived from GL-5-JCK human glioblastoma DNA transfection. The transforming DNA was molecularly cloned by using cosmid vector pJB8 to determine its structure and origin. Analyses of selected clones revealed that the transforming DNA consisted of three portions of human DNA sequences, with the 3' half of the c-raf-1 gene as its middle portion. This raf region was about 20 kilobases long and contained exons 8 to 17 and the poly(A) addition site. RNA blot analysis showed that the raf-related transforming DNA was transcribed into 5.3-, 4.8-, and 2.5-kilobase mRNAs; the 2.5-kilobase transcript was thought to be the major transcript. Immunoprecipitation analyses revealed that a 44-kilodalton raf-related protein was specifically expressed in the NIH 3T3 transformants. The raf-related transforming DNA was considered to be activated when its amino-terminal sequence was truncated and the DNA was coupled with a foreign promoter sequence. On hybridization analysis of the original GL-5-JCK glioblastoma DNA, no rearrangement of c-raf-1 was detectable in the tumor DNA. The rearrangement of c-raf-1 may have occurred during transfection or may have been present in a small population of the original tumor cells as a result of tumor progression.

Original languageEnglish
Pages (from-to)1776-1781
Number of pages6
JournalMolecular and Cellular Biology
Volume7
Issue number5
Publication statusPublished - 1987
Externally publishedYes

Fingerprint

Molecular Cloning
DNA
Genes
Glioblastoma
Transfection
Neoplasms
Cosmids
Poly A
Gene Rearrangement
Immunoprecipitation
Exons
Clone Cells
RNA
Messenger RNA
Population
Proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Fukui, M., Yamamoto, T., Kawai, S., Mitsunobu, F., & Toyoshima, K. (1987). Molecular cloning and characterization of an activated human c-raf-1 gene. Molecular and Cellular Biology, 7(5), 1776-1781.

Molecular cloning and characterization of an activated human c-raf-1 gene. / Fukui, M.; Yamamoto, T.; Kawai, S.; Mitsunobu, Fumihiro; Toyoshima, K.

In: Molecular and Cellular Biology, Vol. 7, No. 5, 1987, p. 1776-1781.

Research output: Contribution to journalArticle

Fukui, M, Yamamoto, T, Kawai, S, Mitsunobu, F & Toyoshima, K 1987, 'Molecular cloning and characterization of an activated human c-raf-1 gene', Molecular and Cellular Biology, vol. 7, no. 5, pp. 1776-1781.
Fukui, M. ; Yamamoto, T. ; Kawai, S. ; Mitsunobu, Fumihiro ; Toyoshima, K. / Molecular cloning and characterization of an activated human c-raf-1 gene. In: Molecular and Cellular Biology. 1987 ; Vol. 7, No. 5. pp. 1776-1781.
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