Protein analysis methods, viral vectors featured in Cold Spring Harbor Protocols|
COLD SPRING HARBOR, N.Y. (Friday, May 1, 2009) – Many proteins do not function by themselves as stand-alone units. Instead, multiple proteins associate to form larger structures called protein complexes. The May issue of Cold Spring Harbor Protocols (www.cshprotocols.org/TOCs/toc5_09.dtl) features a set of methods that can be used to analyze protein complexes. An additional featured article details the generation of viral vectors for gene transfer.
Systematic Monitoring of Protein Complex Composition and Abundance by Blue-Native PAGE, written by Harvey Millar and colleagues from the University of Western Australia (www.biochem.biomedchem.uwa.edu.au/Our_People/home_pages/research_staff/millar), describes multiple experimental approaches using polyacrylamide gel electrophoresis (PAGE). Blue-native PAGE (BN-PAGE) allows a range of protein complexes to be visualized. When combined with sodium dodecyl sulfate PAGE (SDS-PAGE), the procedure can resolve the complexes and their subunits by their molecular weight. In conjunction with differential in-gel electrophoresis (DIGE), BN-PAGE can be used to quantify changes in protein complex abundance or subunit composition between different samples. The article presents detailed methodology for BN-PAGE, SDS-PAGE, and DIGE. It is freely available on the website for Cold Spring Harbor Protocols (cshprotocols.cshlp.org/cgi/content/full/2009/5/pdb.prot5221).
Genetically modified adenoviruses serve as one of the most versatile and efficient gene delivery systems in use today. Laboratories throughout the world use adenoviruses for the delivery of DNA to cells for basic science and for gene therapy applications. Unlike most other vectors, adenoviruses can infect post-mitotic cells, which makes them particularly useful as vectors for gene delivery into cells like neurons. In the May issue of Cold Spring Harbor Protocols, Robin Parks and colleagues from the Ottawa Health Research Institute (www.ohri.ca/profiles/parks_detail.asp) provide Construction and Characterization of Adenovirus Vectors, a set of detailed instructions for the generation, propagation, purification, and characterization of adenovirus vectors. This method is freely accessible on the website for Cold Spring Harbor Protocols (cshprotocols.cshlp.org/cgi/content/full/2009/5/pdb.prot5011).
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Cold Spring Harbor Protocols (www.cshprotocols.org) is a monthly peer-reviewed journal of methods used in a wide range of biology laboratories. It is structured to be highly interactive, with each protocol cross-linked to related methods, descriptive information panels, and illustrative material to maximize the total information available to investigators. Each protocol is clearly presented and designed for easy use at the bench—complete with reagents, equipment, and recipe lists. Life science researchers can access the entire collection via institutional site licenses, and can add their suggestions and comments to further refine the techniques.
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