We have constructed and characterized a Chlamydomonas reinhardtii tota
l genomic library in yeast artificial chromosomes (YACs). The library
contains 7500 clones with inserts ranging in size from 100-200 kb. The
representation of the library was assessed by screening one-third of
it with a probe derived from the dispersed repeat, Gulliver, which occ
urs similar to 13 times in the genome. At least 10 of these Gulliver l
oci were isolated within 15 independent YACs. Two of these YACs encomp
ass the Gulliver element designated G, which was reported to map to th
e uni linkage group (ULG). The end clones of these two YACs have been
genetically mapped by RFLP analysis in an interspecific cross and ther
eby shown to be closely linked to the APM locus on the ULG. A third un
i-specific YAC has also been isolated and its ends have been mapped by
RFLP analysis. Genetic and RFLP analysis of these and other YACs indi
cates that the frequency of chimeric YACs in the library is very low.
The library was constructed in a second generation vector that enables
plasmid rescue of YAC end clones as well as copy number amplification
of artificial chromosomes. We provide evidence that amplification of
intact YACs requires a rad1.rad52 yeast strain.