Convenient transfer of lacZ-gene fusions to phage M13 by in vivo recombination and their use for nucleotide sequencing.
We describe a method that facilitates the sequencing of lacZ fusion joints based on in vivo subcloning onto phage M13. The method is useful for lacZ fusions that are isolated with the transposable lambda placMu phage into plasmids carrying the pBR322 bla gene. In vivo cloning of lacZ fusions is accomplished by recombination with two M13 phages carrying 5' or 3' segments of the bla gene, adjacent but differing in orientation to lacZ'. The presented method allows rapid sequencing of many fusion joints without subcloning in vitro.
Gött
P
P
Department of Biology, University of Konstanz, F.R.G.
Ehrmann
M
M
Boos
W
W
D001435:Bacteriophages / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D004251:DNA Transposable Elements; D007763:Lac Operon; D010957:Plasmids; D011995:Recombination, Genetic
D004251:DNA Transposable Elements