A chromosome integration system for stable gene transfer into Thermus flavus.
We have developed a chromosomal integration system for gene transfer into the extreme thermophile Thermus flavus. The system relies on integration at the site of leuB (3-isopropylmalate dehydrogenase) which was cloned from T. flavus. The leuB gene was insertionally inactivated in vitro with a thermostable kanamycin-resistance gene and transformed in single-copy into the chromosome of T. flavus on a plasmid vector. Gene replacement strains required leucine for growth, were stably kanamycin-resistant and could grow in the presence of kanamycin at temperatures up to 55 degrees C.
Weber
J M
JM
ThermoGen, Inc., Chicago, IL 60612, USA.
Johnson
S P
SP
Vonstein
V
V
Casadaban
M J
MJ
Demirjian
D C
DC
D050539:3-Isopropylmalate Dehydrogenase; D000429:Alcohol Oxidoreductases / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D004355:Drug Stability; D018014:Gene Transfer Techniques; D008040:Genetic Linkage; D006358:Hot Temperature; D007612:Kanamycin / Q000494:pharmacology; D007613:Kanamycin Resistance / Q000235:genetics; D007930:Leucine / Q000378:metabolism; D015183:Restriction Mapping; D013824:Thermus / Q000235:genetics* / Q000254:growth & development
D007612:Kanamycin; D000429:Alcohol Oxidoreductases; D050539:3-Isopropylmalate Dehydrogenase; D007930:Leucine