Open Access

Proteome purification

  • Jonathan B Weitzman
Genome Biology20023:spotlight-20020308-01

DOI: 10.1186/gb-spotlight-20020308-01

Published: 8 March 2002

In the March 5 Proceedings of the National Academy of Sciences, Braun et al. describe a methodology to perform high-throughput purification of human proteins using several affinity-tags (Proc Natl Acad Sci USA2002,99:2654-2659). They exploited the Gateway recombinational cloning system that employs a versatile 'master' vector for easy transfer into different expression vectors. They chose a test set of 32 human genes and expressed them in four different vectors with different affinity tagsL: the His6tag, calmodulin-binding peptide, glutathione-S-transferase or maltose-binding protein. Using different denaturing and nondenaturing purification conditions they were able to isolate around 80% of expressed proteins. Braun et al.demonstrated that the system can be easily scaled-up, and then applied the same approach to a set of 336 human cDNAs with a success rate of around 60%. This approach allows for optimization of protein purification on a proteome-scale.

References

  1. Proceedings of the National Academy of Sciences, [http://www.pnas.org]
  2. Global efforts in structural genomics.
  3. Proteome-scale purification of human proteins from bacteria., [http://www.pnas.org/cgi/content/abstract/99/5/2654]
  4. DNA cloning using in vitro site-specific recombination.

Copyright

© BioMed Central Ltd 2002

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