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

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5' Flanking Region
Transcription
Erythropoietin
Genes
Kidney
Plasmids
RNA-Directed DNA Polymerase
Gels
RNA
In Vitro Techniques

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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In vitro reconstitution of an erythropoietin gene transcription system using its 5′-flanking sequence and a nuclear extract from anemic kidney. / Tsuchiya, Terumasa; Ochiai, Hirokazu; Imajoh-Ohmi, Shinobu; Ueda, Masatsugu; Suda, Toshio; Nakamura, Michio; Kanegasaki, Shiro.

In: Biochemical and Biophysical Research Communications, Vol. 182, No. 1, 15.01.1992, p. 137-143.

Research output: Contribution to journalArticle

Tsuchiya, Terumasa ; Ochiai, Hirokazu ; Imajoh-Ohmi, Shinobu ; Ueda, Masatsugu ; Suda, Toshio ; Nakamura, Michio ; Kanegasaki, Shiro. / In vitro reconstitution of an erythropoietin gene transcription system using its 5′-flanking sequence and a nuclear extract from anemic kidney. In: Biochemical and Biophysical Research Communications. 1992 ; Vol. 182, No. 1. pp. 137-143.
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