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Analysis of Lysin A from Two Novel Mycobacteriophages
Author(s) -
Surendranathan Nandini
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.939.19
Subject(s) - lysin , bacteriophage , mycobacterium smegmatis , peptidoglycan , biology , gene , microbiology and biotechnology , genetics , phage therapy , computational biology , mycobacterium tuberculosis , escherichia coli , tuberculosis , medicine , pathology
As antibiotic resistant bacteria increase in prevalence, bacteriophage have been studied as an alternative to traditional antimicrobials. Through the Phage Hunters project funded by the Howard Hughes Medical Institute (HHMI), students have discovered over fifteen hundred novel bacteriophages that infect Mycobacterium smegmatis (M.Smeg), a bacterium related to Mycobacterium tuberculosis . At Montclair State University, the program has resulted in the isolation and genomic sequencing of numerous phage variants over the past five years. However, while multiple phages have been identified and sequenced, the genes required for their infectivity have not been studied as extensively. In order to gain a better understanding of how these bacteriophage function, this project has focused on the bioinformatic analysis and protein characterization of one gene, Lysin A, in two isolated bacteriophage, ShiVal and TeardropMSU. Lysin A is a protein that enables the virus to break out of the host and has the potential to serve as an antibiotic in its own right. ShiVal is part of the mycobacteriophage cluster B, the second largest group, while TeardropMSU is member of the mycobacteriophage cluster E. Structurally, Lysin A requires a murimidase homology sequence as well as a Zn + binding site and peptidoglycan binding site that enables the enzyme to hydrolyze peptidoglycan. The bioinformatics data indicates that, while some of these critical regions appear in both sequences, there is only a three percent identity between Lysin A of the two bacteriophage which suggests that both lysins could be structurally different from one other. In addition to analyzing sequence comparisons between the two genes, Lysin A was directly isolated from ShiVal and TeardropMSU using PCR. TeardropMSU Lysin A was inserted into a cloning vector to allow for insertion into an expression vector. Protein expression and analysis of these two forms of the protein will allow us to clarify the mechanism of Lysin A activity and determine how the structural differences impact enzyme function. Support or Funding Information I would like to thank the Science Honors Innovation Program (SHIP) and the department of Biology and Molecular Biology at Montclair State University.