In vitro reconstitution of an erythropoietin gene transcription system using its 5′-flanking sequence and a nuclear extract from anemic kidney

Terumasa Tsuchiya, Hirokazu Ochiai, Shinobu Imajoh-Ohmi, Masatsugu Ueda, Toshio Suda, Michio Nakamura, Shiro Kanegasaki

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

We have developed an in vitro transcription system for the erythropoietin (Epo) gene. This system uses a plasmid carrying 0.2 kb of 5′-flanking sequence from the human Epo gene, rNTPs and a nuclear extract from mouse kidney. The transcribed RNA was assayed by primer extension with an end-labeled primer complementary to the sequence of the plasmid, dNTPs and reverse transcriptase. The primer extension product corresponding to the transcript was detected on a sequencing gel. The in vitro promoter activity of the Epo 5′-flanking sequence was observed with a nuclear extract from anemic kidney but not with that from normal kidney.

Original languageEnglish
Pages (from-to)137-143
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume182
Issue number1
DOIs
Publication statusPublished - 1992 Jan 15

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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