Overexpression of gene encoding an ABC transpoter multidrug of Lactobacilllus acidophilus

Abstract

Multidrug resistance (MDR) – type of transporters mediate the active extrusion of structurally and functionally dissimilar compounds from the cells, thereby rendering cells resistance to a range of drugs. ATP- binding cassette (ABC) transporters are characterized as MDR transporters utilizing ATP hydrolysis. LBA0597 gene encoding a multidrug resistance ABC transporter in Lactobacillus acidophilus NCFM has been structurally sequenced but not expressed and functionally unknown for drug resistance mechanism. Consequently, this research was aimed at overexpression and evaluation of this multidrug resistance gene encoding active ABC transporters. The objective was achieved by exploiting the E.coli BL21 (DE3) plysS host expression system and performing disc diffussion method for antibiotic resistance assay. The expressed recombinant protein encoded by the gene was detected by SDS-PAGE at 56 kDa bands as well as showed high resistance to eight kinds of tested antibiotics such as cephalothin (1mg/ml), ciprofloxacin (100 µ g/ml), oxacillin (10mg/ml), vancomycin (1mg/ml), kanamycin (1mg/ml), ampicillin (1mg/ml), erythromycin (100mg/ml), and chloramphenicol (10mg/ml). Our results reported firstly on successful overexpression of multidrug resistance ABC transporter gene of Lactobacillus acidophilus NCFM in E.coli. Lactobacillus acidophilus NCFM lacked the plasmid harboring resistance gene but showed high resistance to tested antibiotics. This strain proved to be a potential probiotic in combination with those antibiotics causing the pseudo membrane colitis in order to prevent the diarrhea.