Cloning of the newt Pleurodeles waltlii chromosomal DNA.
Pleurodeles waltlii genomic DNA has been cloned using several phage lambda vectors. We have isolated approx. 600 000 clones, which correspond to about 20% of the total DNA sequences of this organism. This constitutes the first large gene library of a Urodele. The low yield of cloning was attributable to the abundance of highly repetitive sequences, since recombinations in the bacterial host could lead to the loss of clones. Indeed, the existence of highly repetitive sequences was directly demonstrated by hybridization between recombinants and the total genome, and some of the cloned DNA was found to be unstable. We suggest new methods for cloning the highly repetitive sequences.
Muller
J P
JP
le Maire
M
M
le Hégarat
J C
JC
Randsholt
N
N
D000818:Animals; D010582:Bacteriophage lambda / Q000235:genetics; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005784:Gene Amplification; D005822:Genetic Vectors; D010999:Pleurodeles / Q000235:genetics*; D012091:Repetitive Sequences, Nucleic Acid; D012455:Salamandridae / Q000235:genetics*
D004247:DNA