Amplification and cloning of guanosine 5'-triphosphate, 3'- diphosphate phosphohydrolase enzyme from Mycobacterium tuberculosis

Mycobacterium tuberculosis, the etiological agent of human tuberculosis (TB), is one of the most successful bacterial pathogens, remaining a leading cause of morbidity and mortality. One-third of the world’s population is infected with M. tuberculosis in a condition known as latency (without clinical manifestations) which facilitates transmission and complicates treatment. Thus, there is a continuous necessity of studies on mycobacterial metabolism. Guanosine 5’-triphosphate, 3’-diphosphate phosphohydrolase from Mycobacterium tuberculosis (MtGpp) is a conserved hypothetical protein similar to Gpp from Escherichia coli and is a potential target for rational drugs design, since it may participate in the guanosine 5’-diphosphate, 3’-diphosphate (ppGpp) biosynthesis, an important regulatory nucleotide for prokaryotic organisms. In this work, we report amplification, cloning, subcloning and overexpression for a Gpp from M. tuberculosis H37Rv. This proves the knowledge about enzymes involved in Mycobacterium metabolism to pave the way the design of new antituberculosis agents and vaccines development.