1. Characterization of a (pckA) mutant of the non-nodulating bacterium Rhizobium pusense NRCPB10 induced by transposon Tn5 mutagenesis
- Author
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P. K. Chakrabartty and Subrata K. Das
- Subjects
Cloning ,chemistry.chemical_classification ,Transposable element ,Genetics ,PckA ,Hypothetical protein ,Mutant ,Tn5 transposon mutagenesis ,Mutagenesis (molecular biology technique) ,Environmental Science (miscellaneous) ,Biology ,Agricultural and Biological Sciences (miscellaneous) ,Amino acid ,Rhizobium pusense ,chemistry ,Original Article ,Phosphoenolpyruvate carboxykinase ,Gene ,Biotechnology - Abstract
Phosphoenolpyruvate carboxykinase (PCK) catalyzes the decarboxylation and phosphorylation of oxaloacetate to phosphoenolpyruvate in the gluconeogenic pathway in most organisms. A pckA gene encoding PCK was cloned and sequenced from strain Rhizobium pusense NRCPB100, a spontaneous rifampicin resistant mutant of R. pusense NRCPB10T (JCM16209T) of a recently described new species of a non-nodulating and non-tumorigenic bacterium. The mapping of the pck gene region following Tn5 mutagenesis located the gene downstream of a transcriptional regulatory protein gene (chvI) and upstream of a conserved hypothetical protein gene. The pck of 1,611 bp was deduced to encode 536 amino acids and showed high homology to the genes of known ATP-dependent PCK enzymes. Phylogenetic analysis of the gene placed it in a cluster with pck of other known members of Rhizobiales. Amino acid sequences of the putative functional regions of the deduced enzyme were found to be conserved.
- Published
- 2012