The effects of temperature on the expression of recombinant phospholipase A₂ in escherichia coli / Mohd 'Izzat Idris

The aim of the research is to study the effects of temperature on the expression of recombinant Pla, in E. coli. There were two clones of E. coli used in this study. Both clones were incubated at 27°C and 37°C respectively. The method of this study was started with 100 µL overnight cultured was transferred into LB broth. After two hours, the cultures were supplemented with 0.2 % arabinose to induce the expression of the PLA2 and incubated at two different temperatures. Samples (PLA2 protein) that have been collected were analysed through SDS-PAGE, Bradford protein quantitation and sPLA2 protein activity. Prior to performing analysis study, the protein must be purified from the host. Purification was done using Maxwell 16 polyhistidine protein purification kit. The finding shows that clone pBAD Thio­ TOPO Pla2 (-TAG 5) and pBAD Thio-TOPO Pla2 (-TAG 8) produced PLA2 protein. Result showed that 37 °C incubation temperature produced higher protein amount when compared to 27 °C incubation temperature. It can be concluded that temperature was found to give effect towards the amount of protein being expressed.