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Whole Genome Sequence and Development of a Transposon Mutagenesis System for Pseudomonas Putida sp.12

Nowadays, Heavy metal pollution is one of the most important environmental problems. All metals 
occurring in the biosphere originated in the earth’s crust; however, the toxic metal pollution of the biosphere 
has intensified rapidly since the onset of the industrial revolution, posing major environmental and 
health problems. Several studies have demonstrated the effect of metals on microbial diversity, biomass 
and activity. It is believed that metal Microbial activity is responsible for the bioremediation of the third 
elements in the periodic table. Selenium is an important trace element that required for in trace amounts for 
growth and metabolism but toxic at elevated concentration. The ability of a single mutant Pseudomonas 
putida sp.12(mariner transposon) has been used to search for genes involved in the biotransformation of 
selenite to red elemental selenium. This study aims to develop a transposon mutagenesis system for the 
model P. putida sp.12 to investigate the gene(s) that involved in selenite reduction. The conjugative plasmid 
pSAM_R1 containing the mariner transposon was used for transposon mutagenesis. A single mutant reliably 
impaired in selenite transformation, found that the mutant clones, contained an inactivation within pmoB, 
one of two copies of the gene encoding the largest subunit of particulate selenite-reduction. Physiological 
analysis of this “white mutant” indicated that the selenite-reducing activity, which was located primarily in 
the cytoplasm of the cells, could be rescued by adding format as an alternative source of electrons