Recombinant MS2 L Is Biologically Functional

Abstract

The single stranded RNA bacteriophage MS2 attacks its host, Escherichia coli, with the help of its lysis protein, L, rather than lytic enzymes. The L protein consists of 75 amino acids, with a hydrophilic N‐terminal that contains mainly basic residues and a hydrophobic C‐terminal end. Prior studies have shown that the L protein is necessary for the cell to lyse, however it is not known if L is sufficient for cell lysis. This study may lead to identifying whether or not the L protein in MS2 is the singular factor in cell lysis. Tests have been conducted using a previously engineered plasmid that contains MS2 L under the control of the arabinose inducible pBAD promoter. Transformed E. coli cultures were induced with arabinose and compared via spectrophotometry. The results show a decrease in the amount of cells within 40 minutes after induction compared with non‐induced and GFP producing controls demonstrating that the L protein is indeed functional and triggering E. coli lysis. The question still remains as to whether or not L is functioning independently or in concert with other E. coli proteins. In order to explore this possibility, a GST‐tag will be cloned in‐frame with the L ORF. After determining whether the GST‐L is biologically functional, we will perform pull‐down experiments to determine whether host proteins are associated with L.Support or Funding InformationThis work was funded by the Hartwick College Chemistry Department as well as a Hartwick College Faculty Research Grant awarded to AJP.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.