Perpustakaan Digital ITB

Terpenoids are natural compounds with various pharmacological activities. The use of Bacillus subtils for terpenoid synthesis through the mevalonate pathway (MVA) has been developed. However, the pathway is still inefficient. Acetoacetyl-coA thiolase, encoded by mmgA gene, is suspected to interfere with the pathway and might be responsible in degrading the main precursor of the MVA pathway. This study aims to design primer for the detection of mmgA and gyrA genes (reference genes) in silico and validated by primer specificity and efficiency tests. This study used B. subtilis Wild Type 168 (WT), MVA6 (WT containing plasmid pDR111-atoB-mvaA-mvaS), B. subtilis mutant BKK24170 (????mmgA::kan), and BKK24170-MVA6 (containing plasmid pDR111-atoB-mvaA-mvaS). The four bacteria were grown and induced to be followed by RNA isolation and qPCR using selected primers. Transformation of plasmid pDR111-atoB-mvaA-mwaS was successfully carried out on BKK24170. RNA was successfully isolated from the four strains. The mmgA and gyrA primers were successfully designed in silico but did not have good efficiency and specificity hence, mmgA mRNA expression analysis could not be carried out.