An improved retroviral vector for assaying promoter activity. Analysis of promoter interference in pIP211 vector

Kazunori Nakajima, Kazuhiro Ikenaka, Kensuke Nakahira, Noriyuki Morita, Katsuhiko Mikoshiba

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

We recently developed a novel promoter assay system using a retroviral vector (pIP200 series). Transcription from the internal promoter, which had been inserted for the promoter assay, was shown to be interfered with by transcription from the upstream long terminal repeat (LTR). Here we report a new high-titer 'self-inactivating' vector, in which transcription interference was virtually eliminated. This new vector was constructed by introducing only a very minor mutation into the 'TATA box' in the 3'-LTR. This mutation was successfully transferred to the 5'-LTR after reverse transcription, yielding a provirus incapable of transcribing viral RNA. The viral titer was not reduced by the mutation, permitting general application of this virus.

Original languageEnglish
Pages (from-to)129-133
Number of pages5
JournalFEBS Letters
Volume315
Issue number2
DOIs
Publication statusPublished - 1993 Jan 4

Fingerprint

Terminal Repeat Sequences
Transcription
Mutation
Proviruses
Assays
TATA Box
Viral RNA
Reverse Transcription
Viruses

Keywords

  • Fluorescein-di-β-d-galactopyranoside
  • Promoter interference
  • Retroviral vector

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

An improved retroviral vector for assaying promoter activity. Analysis of promoter interference in pIP211 vector. / Nakajima, Kazunori; Ikenaka, Kazuhiro; Nakahira, Kensuke; Morita, Noriyuki; Mikoshiba, Katsuhiko.

In: FEBS Letters, Vol. 315, No. 2, 04.01.1993, p. 129-133.

Research output: Contribution to journalArticle

Nakajima, Kazunori ; Ikenaka, Kazuhiro ; Nakahira, Kensuke ; Morita, Noriyuki ; Mikoshiba, Katsuhiko. / An improved retroviral vector for assaying promoter activity. Analysis of promoter interference in pIP211 vector. In: FEBS Letters. 1993 ; Vol. 315, No. 2. pp. 129-133.
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