One-step cloning and chromosomal integration of DNA.
We describe "clonetegration", a method for integrating DNA into prokaryotic chromosomes that approaches the simplicity of cloning DNA within extrachromosomal vectors. Compared to existing techniques, clonetegration drastically decreases the time and effort needed for integration of single or multiple DNA fragments. Additionally, clonetegration facilitates cloning and expression of genetic elements that are impossible to propagate within typical multicopy plasmids.
Department of Bioengineering and ‡Department of Pediatrics, Stanford University , California 94305, United States.
François
F
St-Pierre
Lun
L
Cui
David G
DG
Priest
Drew
D
Endy
Ian B
IB
Dodd
Keith E
KE
Shearwin
D002876:Chromosomes, Bacterial / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA / Q000235:genetics*; D017931:DNA Primers; D004926:Escherichia coli / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods; D005822:Genetic Vectors; D012486:Salmonella typhimurium / Q000235:genetics*; D014170:Transformation, Genetic
D017931:DNA Primers; D004247:DNA