A general cloning system to selectively isolate any eukaryotic or prokaryotic genomic region in yeast.

ABSTRACT:

Background

Transformation-associated recombination (TAR) cloning in yeast is a unique method for selective isolation of large chromosomal fragments or entire genes from complex genomes. The technique involves homologous recombination, during yeast spheroplast transformation, between genomic DNA and a TAR vector that has short (approximately 60 bp) 5' and 3' gene targeting sequences (hooks).

Result

TAR cloning requires that the cloned DNA fragment carry at least one autonomously replicating sequence (ARS) that can function as the origin of replication in yeast, which prevents wide application of the method. In this paper, we describe a novel TAR cloning system that allows isolation of genomic regions lacking yeast ARS-like sequences. ARS is inserted into the TAR vector along

SUBMITTER: Noskov VN 

PROVIDER: S-EPMC156606 | biostudies-literature | 2003 Apr

REPOSITORIES: biostudies-literature