PMID: 8972855Dec 1, 1996Paper

A shuttle system for transfer of YACs between yeast and mammalian cells

Nucleic Acids Research
K Simpson, C Huxley

Abstract

The development of a system for shuttling DNA cloned as yeast artificial chromosomes (YACs) between yeast and mammalian cells requires that the DNA is maintained as extrachromosomal elements in both cell types. We have recently shown that circular YACs carrying the Epstein-Barr virus origin of plasmid replication (oriP) are maintained as stable, episomal elements in a human kidney cell line constitutively expressing the viral transactivator protein EBNA-1. Here, we demonstrate that a 90-kb episomal YAC can be isolated intact from human cells by a simple alkaline lysis procedure and shuttled back into Saccharomyces cerevisiae by spheroplast transformation. In addition, we demonstrate that the 90-kb YAC can be isolated intact from yeast cells. The ability to shuttle large, intact fragments of DNA between yeast and human cells should provide a powerful tool in the manipulation and analysis of functional regions of mammalian DNA.

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Citations

Jul 17, 2008·Molecular Therapy : the Journal of the American Society of Gene Therapy·Michele M P LufinoRichard Wade-Martins
Apr 1, 1997·Nature Genetics·M A Rosenfeld
Mar 9, 2002·Journal of Virology·Peter C AngelettiPaul F Lambert
Oct 29, 2005·Journal of Virology·Lei Chen, Michael Lagunoff
Mar 16, 2005·Journal of Virology·Nazmul AhsanKenzo Takada
Jun 6, 2000·Journal of Obstetric, Gynecologic, and Neonatal Nursing : JOGNN·J V Schmidt, P R McCartney
Nov 19, 2011·Bioengineered Bugs·Wing CheungClare Huxley
Dec 17, 2016·Nucleic Acids Research·David M BrownJohn I Glass
Jan 19, 2020·Experimental Cell Research·David M Brown, John I Glass
Jul 1, 1997·Genetic Analysis : Biomolecular Engineering·E FrengenP J de Jong

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