Rapid modification of bacterial artificial chromosomes by ET-recombination.
We present a method to modify bacterial artificial chromosomes (BACs) resident in their host strain. The method is based on homologous recombination by ET-cloning. We have successfully modified BACs at two distinct loci by recombination with a PCR product containing homology arms of 50 nt. The procedure we describe here is rapid, was found to work with high efficiency and should be applicable to any BAC modification desired.
Gene Expression Program, European Molecular Biology Laboratory, Meyerhofstrasse 1, D-69117 Heidelberg, Germany.
J P
JP
Muyrers
Y
Y
Zhang
G
G
Testa
A F
AF
Stewart
D001426:Bacterial Proteins / Q000378:metabolism*; D002876:Chromosomes, Bacterial / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods*; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D005090:Exodeoxyribonucleases / Q000378:metabolism*; D008957:Models, Genetic; D011995:Recombination, Genetic
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C084497:RecT protein, E coli; D005090:Exodeoxyribonucleases; C038225:recE protein, E coli