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

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

DOI:
10.3791/55576-v

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12:23 min

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May 16, 2017

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, ,
1Division of Biological Science and Technology,Yonsei University

Chapitres

  • 00:05Titre
  • 00:55Recombinant Protein Production and Purification of rBC1531
  • 04:47Removal of Affinity Tags by Thrombin Proteolysis
  • 06:32Crystallization Screening and Optimization of rBC1531
  • 07:30Colorimetric NTPase Assay
  • 08:33Results: Purification, Characterization, and Crystallization of rBC1531
  • 10:31Conclusion

Summary

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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.

Tags

Recombinant ProteinBacillusNucleotide PyrophosphorylaseBcMazGProtein ExpressionProtein CrystallizationBiophysical StudiesE. ColiChemically Competent CellsTransformationPlasmid DNADNA SequencingProtein PurificationIPTG InductionCell LysisProtein Characterization

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