Identification of DNA sequences required for transcription of the human alpha 1-globin gene in a new SV40 host-vector system

Cell. 1981 Dec;27(2 Pt 1):279-88. doi: 10.1016/0092-8674(81)90411-6.

Abstract

We have developed a rapid and simple method for studying the transcription of cloned eucaryotic genes, which involves transfecting SV40-transformed monkey cell lines (COS cells) with derivatives of the plasmid pBR322 that contain the SV40 viral replication origin but lack regions necessary for viral transcription (SV-ORI vectors). Because COS cells produce SV40 T antigen and are permissive for SV40 viral replication, transfected SV-ORI plasmids replicate to a high copy number. SV-ORI plasmids carrying a human alpha-globin gene are also replicated in COS cells. Moreover, the alpha-globin gene is faithfully transcribed to produce high levels of RNA, which is accurately processed to produce authentic alpha-globin mRNA. We have used this transcription system to demonstrate that a sequence located between 55 and 87 base pairs upstream from the mRNA capping site is required for efficient transcription of the alpha-globin gene in COS cells.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • Cell Line
  • Chlorocebus aethiops
  • DNA, Recombinant
  • Genetic Vectors
  • Globins / genetics*
  • Humans
  • Kidney
  • Operon
  • Simian virus 40 / genetics
  • Transcription, Genetic*
  • Transfection

Substances

  • DNA, Recombinant
  • Globins