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

Gene
P J RipollB Arveiler

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.

References

Aug 1, 1992·Proceedings of the National Academy of Sciences of the United States of America·R A McKnightL Hennighausen
May 1, 1990·Genetic Analysis, Techniques and Applications·J LundebergM Uhlén
Oct 12, 1990·Science·M Koob, W Szybalski
Jun 1, 1987·Molecular & General Genetics : MGG·K NishiwakiY Oshima
Jan 1, 1994·Trends in Genetics : TIG·J PetitB Arveiler
Jan 1, 1993·Mammalian Genome : Official Journal of the International Mammalian Genome Society·D R SmithD T Moir

❮ Previous
Next ❯

Citations

Nov 26, 2003·FEMS Microbiology Reviews·Natalay Kouprina, Vladimir Larionov

❮ Previous
Next ❯

Related Concepts

Related Feeds

Artificial Chromosomes

Artificial chromosomes are genetically engineered chromosomes derived from the DNA of a species. Discover the latest research on artificial chromosomes here.