Email the Director
John Leigh, Ph.D.
Markerless Mutagenesis Procedure
Currently we have two systems for producing markerless mutations in Methanococus maripaludis. Our original system, using pCRPrtNeo to make mutations in strain Mm900, takes advantage of the susceptibility to the purine analog 8-azahypoxanthine conferred by the hpt (hypoxanthine / guanine
phosphoribosyltransferase) gene. More recently we developed pCRUptNeo for markerless mutagenesis of strain Mm901. This system is what we now use routinely and takes advantage of the susceptibility to the pyrimidine analog 6-azauaracil conferred by the upt (uracil
phosphoribosyltransferase) gene. We also developed a method that uses pBlPrt to stably incorporate
constructs into the genome, at the site of the upt gene (see markerless insertions procedure).
|Markerless Mutagenesis Procedure with pCRUptNeo and Mm901
|Markerless Mutagenesis Procedure with pCRPrtNeo and Mm900