Group II introns as controllable gene targeting vectors for genetic manipulation of bacteria.
Mobile group II introns can be retargeted to insert into virtually any desired DNA target. Here we show that retargeted group II introns can be used for highly specific chromosomal gene disruption in Escherichia coli and other bacteria at frequencies of 0.1-22%. Furthermore, the introns can be used to introduce targeted chromosomal breaks, which can be repaired by transformation with a homologous DNA fragment, enabling the introduction of point mutations. Because of their wide host range, mobile group II introns should be useful for genetic engineering and functional genomics in a wide variety of bacteria.
Karberg
M
M
Institute for Cellular and Molecular Biology, Department of Chemistry and Biochemistry, School of Biological Sciences, University of Texas at Austin, Austin, TX 78712, USA.
Guo
H
H
Zhong
J
J
Coon
R
R
Perutka
J
J
Lambowitz
A M
AM
D001483:Base Sequence; D015139:Blotting, Southern; D002875:Chromosomes / Q000235:genetics; D018388:Codon, Terminator; D004926:Escherichia coli / Q000235:genetics*; D015723:Gene Library; D018014:Gene Transfer Techniques; D005798:Genes, Bacterial; D005818:Genetic Engineering; D005822:Genetic Vectors; D007438:Introns; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic
D018388:Codon, Terminator