Abstract
We report here the first use of directed mutagenesis in Methanosarcina acetivorans C2A. The method employs homologous recombination-mediated gene replacement and was used to construct a variety of proline auxotrophs with mutations in the proABC locus. Each mutation was also complemented in trans with autonomously replicating Methanosarcina-Escherichia plasmid shuttle vectors.
Publication types
-
Evaluation Study
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Aldehyde Oxidoreductases / genetics
-
Archaeal Proteins / genetics*
-
Archaeal Proteins / metabolism
-
Cloning, Molecular
-
Genes, Archaeal
-
Genetic Complementation Test / methods*
-
Glutamate-5-Semialdehyde Dehydrogenase
-
Methanosarcina / genetics*
-
Methanosarcina / metabolism
-
Molecular Sequence Data
-
Mutagenesis, Insertional / methods*
-
Phosphotransferases (Carboxyl Group Acceptor) / genetics
-
Plasmids / genetics*
-
Proline / biosynthesis*
-
Pyrroline Carboxylate Reductases / genetics
-
Recombination, Genetic
-
Sequence Analysis, DNA
Substances
-
Archaeal Proteins
-
Proline
-
Aldehyde Oxidoreductases
-
Glutamate-5-Semialdehyde Dehydrogenase
-
Pyrroline Carboxylate Reductases
-
Phosphotransferases (Carboxyl Group Acceptor)
-
glutamate 5-kinase