Will ß-lactams Save You or Will You Meet Certain Death?

Abstract

The purpose of this experiment was to determine which gene or genes in Elizabethkingiameningoseptica encode for the enzyme Beta-lactamase. E. meningoseptica was originally described as FlavobacteriumMeningosepticum in 1959 by American bacteriologist Elizabeth O. King. It is ubiquitously found in water and soil, but because of its survival in hospital environments, outbreaks can occur due to exposure to a contaminated water source or medical devices (1). This bacterium /pathogen is resistant to multiple antimicrobials commonly used for bacteria and conventional empirical antimicrobials targeting those organisms may result in unfavorable outcomes (5).

        At the beginning of the experiment we were given a DNA strand, E.ano_Ag1_bla3134|putative MBL,  that contained 16-30 bases.  After that we designed a forward and a reverse primer in order to use the PCR method. Through this method we generated thousands to millions of copies of the gene we were assigned. It was then dyed for gel electrophoresis, which indicated how many bases were in our gene sequence. Our sample corresponded with 639 bases on the molecular weight standard, it was also the amount we counted ourselves. The ligation reaction and heat shock was used to transform the plasmid into E. Coli along with gene that is known to encode for Kanamycin resistance, so as to easily test if transformation occurred. The next few steps after that were done by our professor and TA’s because E. meningosepticais classified as level II by the CDC and we therefore did not have the training to work with it directly. We, group 23 concluded our research with the finding that our starting DNA strand was indeed a beta-lactamase but that it had a low capacity of resistance for the antibiotic that we were testing it for.