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Investigation of Controlled Expression of MS2 Lysis Protein
Author(s) -
Rasefske Kelsey Anne,
Piefer Andrew J.
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.622.8
Subject(s) - lysis , bacteriophage ms2 , escherichia coli , biology , bacteriophage , operon , rna , coliphage , gene , bacteria , microbiology and biotechnology , biochemistry , genetics
Bacteriophage are a class of virus that exclusively infect bacteria cells, with MS2 a group 1 RNA coliphage (affecting the bacteria Escherichia coli). The viral life cycle is comprised of six stages, the first two involving attachment and injection of the RNA genome directly into the host. Lysis is the final stage; including compromise of host cell membrane integrity and release of progeny phage, it is mediated by the lysis, or “L” protein. Consisting of seventy‐five amino acids,(1) L is one of four proteins coded for by the simple MS2 genome, approximately 3500 nucleotides in length.(2) It produces a larger hole in the host cell membrane compared to other bacteriophage lysis proteins and L retains lysis ability, although less efficient, in truncated form.(1) The virus‐free expression of L and the lysis process it elicits is the focus of this study. Toward this goal, a synthetic L‐gene was cloned and ligated to an inducible arabinose operon and RBS using “Bio‐brick” technology. After induction, lysis will be induced and relativity cell density monitored over time via spectrophotometry. A C‐terminally‐fused L‐GFP construct, to track protein localization within the cell and to provide a “handle” for future pull‐down experiments, will also be constructed. Investigation into the role of bacterial host proteins involved in the lysis process has not been previously reported. Support or Funding Information This study was graciously funded by Drs. Allen and Judy Freedman. (Hartwick College Freedman Prize ‐ Physical and Life Sciences)