Construction of a mouse yeast artificial chromosome library in a recombination-deficient strain of yeast.
We have constructed a new generation yeast artificial chromosome (YAC) library from female C57BL/10 mice in a recombination-deficient strain of Saccharomyces cerevisiae carrying a mutation in the RAD52 gene. The YAC library contains 41,568 clones with an average insert size of 240 kilobases, representing a greater than threefold coverage of the mouse genome. Currently, the library can be screened by polymerase chain reaction and we have isolated positive clones at a number of loci in the mouse genome. This rad52 library should enable a long-term assessment of the effect of one of the yeast recombination pathway genes on both, genome-wide YAC clone stability and the frequency of chimaeric clones.
Chartier
F L
FL
Department of Biochemistry and Molecular Genetics, St Mary's Hospital Medical School, London, UK.
Keer
J T
JT
Sutcliffe
M J
MJ
Henriques
D A
DA
Mileham
P
P
Brown
S D
SD
D000818:Animals; D003001:Cloning, Molecular / Q000379:methods*; D005260:Female; D015723:Gene Library; D051379:Mice; D008810:Mice, Inbred C57BL; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014170:Transformation, Genetic