Recombinant Protein Expression, Crystallization, and Biophysical Studies of a Bacillus-conserved Nucleotide Pyrophosphorylase, BcMazG

J Vis Exp. 2017 May 16:(123):55576. doi: 10.3791/55576.

Abstract

To overcome safety restrictions and regulations when studying genes and proteins from true pathogens, their homologues can be studied. Bacillus anthracis is an obligate pathogen that causes fatal inhalational anthrax. Bacillus cereus is considered a useful model for studying B. anthracis due to its close evolutionary relationship. The gene cluster ba1554 - ba1558 of B. anthracis is highly conserved with the bc1531- bc1535 cluster in B. cereus, as well as with the bt1364-bt1368 cluster in Bacillus thuringiensis, indicating the critical role of the associated genes in the Bacillus genus. This manuscript describes methods to prepare and characterize a protein product of the first gene (ba1554) from the gene cluster in B. anthracis using a recombinant protein of its ortholog in B. cereus, bc1531.

Publication types

  • Video-Audio Media

MeSH terms

  • Bacillus anthracis / genetics*
  • Bacillus cereus / growth & development*
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Crystallization
  • Multigene Family
  • Pyrophosphatases / chemistry
  • Pyrophosphatases / genetics*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics*

Substances

  • Bacterial Proteins
  • Recombinant Proteins
  • Pyrophosphatases
  • nucleotide pyrophosphatase