In vitro transcription of c-jun gene using fractionated nuclear extract from regenerating rat liver.

Dipali Sharma, Shailesh Kumar Choudhary, Aparna Dixit

Research output: Contribution to journalArticlepeer-review

Abstract

Regenerating rat liver serves as a source of proliferating cells, such a system can be used to study the regulation of genes involved in proliferation. We have established an in vitro transcription capable of supporting accurate transcription of cloned c-jun gene using fractionated nuclear extract prepared from partially hepatectomized rat liver. EcoR I linearized plasmid -132/+170 jun-CAT containing c-jun gene promoter region (-132 to +170 nucleotides) was transcribed in an in vitro run-off transcription assay and a transcript of expected size i.e. 370 nucleotides was obtained. The intensity of the transcript was dependent on the concentration of fractionated nuclear extract as well as template. The transcription was α-amanitin sensitive indicating that it is directed by RNA polymerase II.

Original languageEnglish (US)
Pages (from-to)1175-1185
Number of pages11
JournalBiochemistry and Molecular Biology International
Volume44
Issue number6
StatePublished - May 1 1998
Externally publishedYes

Keywords

  • In vitro transcription
  • Regenerating rat liver
  • c-jun gene

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Genetics

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