Recombinant Protein Production and Purification of rBC1531
4:47
Removal of Affinity Tags by Thrombin Proteolysis
6:32
Crystallization Screening and Optimization of rBC1531
7:30
Colorimetric NTPase Assay
8:33
Results: Purification, Characterization, and Crystallization of rBC1531
10:31
Conclusion
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The overall goal of this procedure is to prepare recombinant protein from non pathogenic bacteria to facilitate biophysical chemical studies such as the protein x-ray crystallography or enzymatic kinetics. This method can help answer key questions
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Bacillus anthracis is the obligate pathogen of fatal inhalational anthrax, so studies of its genes and proteins are strictly regulated. An alternative approach is to study orthologous genes. We describe biophysicochemical studies of a B. anthracis ortholog in B. cereus, bc1531, requiring minimal experimental equipment and lacking serious safety concerns.