Pengklonan kandidat gen resisten terhadap selenium dari isolat bakteri termofilik Geobacillus sp20k

Abstract

Termophilic bacteria which is resistant to selenium has a specific mechanism enabling it to accumulate and make the toxic selenium becomes nontoxic to the bacteria itself. The mechanism of action is believed because of the existence of a gene that function to confer resistance to Se (Selenium). However, the existence of the gene itself has not been scientifically proven. Therefore it is necessary to do research on thermophilic bacteria to obtain isolate of thermophilic bacteria from hot springs that have gene encoding resistance to selenium (Se) and clone the gene into an E. coli. Isolated genomic DNA of the thermophilic bacteria as well as the plasmid were cut with Eco RI enzyme. Fragments of bacterial DNA was ligated into the vector and then transformed into E. coli cells. Transformants were tested by using blue white colony to show the growth of white colonies indicated the presence of inserted DNA fragment in pGEM3zf while the growth of blue colonies indicated the absence of DNA fragments inserted in pGEM3zf. Molecular weight of the recombinant plasmid was measured using standard curve with obtained assistance line following equation: Y = 19473.807-6294.596X; where X = distance migration and Y = base pairs. Molecular weight of the recombinant plasmid obtained was 14 438 bp.