Molecular cloning of bacterial DNA in vivo using a transposable R6K ori and a P1vir phage.
A new method of cloning in vivo using the P1vir phage and transposon Tn5-rpsL oriR6K was developed. The method relies upon recircularization of transducing DNA containing a transposon insertion in a recombination-deficient strain of Escherichia coli K-12 and subsequent stable replication of the recircularized DNA. Using this method, we were able to clone in vivo the chromosomal region located between approximately 7.1 and 9.2 min on the E. coli K-12 map in a 95-kb plasmid.
Stojiljkovic
I
I
Rugjer Boskovic Institute, Zagreb, Croatia.
Bozja
J
J
Salaj-Smic
E
E
D017101:Bacteriophage P1 / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods*; D004261:DNA Replication / Q000235:genetics; D004251:DNA Transposable Elements / Q000235:genetics*; D004270:DNA, Circular; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D011995:Recombination, Genetic / Q000235:genetics; D000091243:Ribosomal Protein S9; D014161:Transduction, Genetic
D004251:DNA Transposable Elements; D004270:DNA, Circular; D029968:Escherichia coli Proteins; D000091243:Ribosomal Protein S9; C000717190:RpsI protein, E coli