Specific isolation of human rDNA genes by TAR cloning.
Selective cloning of human DNA in YACs from monochromosomal human/rodent hybrid cells lines and radiation hybrids can be accomplished by transformation-associated recombination (TAR) between Alu-containing vector(s) and human DNA in yeast. We have expanded this approach to the specific isolation of repetitive genes from the human genome. Highly selective isolation of human rDNA was accomplished using total human DNA and a pair of differentially marked linear TAR cloning vectors where one contained a small fragment of a human rDNA repeat and the other had an Alu repeat as targeting sequences. About half the transformants that acquired both vectors markers had YACs with human rDNA inserts. These results suggest that TAR can be applied to the general isolation of gene families and amplified region from genomic DNAs.
Kouprina
N
N
Laboratory of Molecular Genetics, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, USA.
Graves
J
J
Cancilla
M R
MR
Resnick
M A
MA
Larionov
V
V
D018244:Chromosomes, Artificial, Yeast / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods*; D004275:DNA, Ribosomal / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014170:Transformation, Genetic
D004275:DNA, Ribosomal