A new yeast artificial chromosome vector designed for gene transfer into mammalian cells

P J Ripoll, A Cowper, S Salmeron, P Dickinson, D Porteous, B Arveiler

Research output: Contribution to journalArticlepeer-review

Abstract

This report describes the construction of a new yeast artificial chromosome (YAC) vector designed for gene transfer into mammalian cells. For ease of use, the two arms of the vector were cloned separately. The vector harbours the Neo and Hyg genes for dominant selection in mammalian cells, a putative human origin of replication, a synthetic matrix attachment region and two loxP sites (one on each arm). The cloning ability of the vector was demonstrated by successful propagation of the cDNA of the cystic fibrosis gene, CFTR, as a YAC in Saccharomyces cerevisiae. A YAC containing the entire CFTR gene was also constructed by retrofitting the two arms of a pre-existing clone (37AB12) with the two arms of the novel vector. Both the cDNA and entire gene containing YACs were circularized in yeast by inducible expression of the Cre recombinase. Recombination occurred very specifically at the loxP sequences present on the two arms of the YAC. Applications of the vector to gene transfer are discussed.
Original languageEnglish
Pages (from-to)163-72
Number of pages10
JournalGene
Volume210
Issue number1
DOIs
Publication statusPublished - 1998

Keywords

  • Animals
  • Base Sequence
  • Cells, Cultured
  • Chromosomes, Artificial, Yeast
  • Cloning, Molecular
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • DNA, Circular
  • DNA, Complementary
  • Gene Expression Regulation, Enzymologic
  • Gene Transfer Techniques
  • Genetic Vectors
  • Integrases
  • Mammals
  • Molecular Sequence Data
  • Saccharomyces cerevisiae
  • Viral Proteins

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