SPECIFIC ISOLATION OF HUMAN RDNA GENES BY TAR CLONING

Selective cloning of human DNA in YACs from monochromosomal human/rode nt hybrid cells lines and radiation hybrids can be accomplished by tra nsformation-associated recombination (TAR) between Alu-containing vect or(s) and human DNA in yeast. We have expanded this approach to the sp ecific isolation of repetitive genes from the human genome. Highly sel ective isolation of human rDNA was accomplished using total human DNA and a pair of differentially marked linear TAR cloning vectors where o ne contained a small fragment of a human rDNA repeat and the other had an Alu repeat as targeting sequences. About half the transformants th at acquired both vectors markers had YACs with human rDNA inserts. The se results suggest that TAR can be applied to the general isolation of gene families and amplified region from genomic DNAs. (C) 1997 Elsevi er Science B.V.