Methods For Visualizing Protein Dynamics Featured In Cold Spring Harbor Protocols

Main Category: Genetics
Also Included In: Biology / Biochemistry
Article Date: 03 Jan 2008 - 3:00 PDT

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This month's release of Cold Spring Harbor Protocols highlights methods that permit scientists to observe protein dynamics in chromosomes and embryos. These methods are freely accessible online (http://www.cshprotocols.org/).

The first protocol, available here, describes how to insert multiple copies of the bacterial lac operator sequence into the chromosomes of mammalian cells. The protocol includes strategies for stabilizing the repeats and for isolating specific cell clones that are tagged at the chromosomal sites of interest.

Once the tagged cell lines have been established, the chromosomal regions of interest can be visualized with a fluorescently labeled Lac repressor protein, which binds to the lac operator sequence. This allows the scientists to examine the structure of the chromatin and to observe the activity of the chromosomes during replication and transcription.

The protocol was developed by Dr. Andrew Belmont's group from the University of Illinois at Urbana-Champaign (http://www.life.uiuc.edu/belmont/). Belmont's lab and others have used this technique to investigate the structure and dynamics of chromatin in live cell cultures, yeast, and various multicellular organisms, including C. elegans, Drosophila, and Arabidopsis.

The second featured method in the current issue of Cold Spring Harbor Protocols describes how to perform immunohistochemistry in whole mouse embryos, which allows scientists to examine the three-dimensional distribution of a protein during specific stages of development. It can be used to detect endogenous proteins as well as the products of transgenes.

The immunohistochemistry protocol was derived from methods used to examine embryonic development in other species, including Xenopus and Drosophila. It describes several tricks to chemically and physically manipulate the embryos so that the antibodies - which detect the protein of interest - can efficiently penetrate the embryonic tissue.

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Article adapted by Medical News Today from original press release.
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It is freely available here.

About Cold Spring Harbor Protocols:

Cold Spring Harbor Protocols (http://www.cshprotocols.org/) is an online resource 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.

About Cold Spring Harbor Laboratory Press:

Cold Spring Harbor Laboratory Press is an internationally renowned publisher of books, journals, and electronic media, located on Long Island, New York. Since 1933, it has furthered the advance and spread of scientific knowledge in all areas of genetics and molecular biology, including cancer biology, plant science, bioinformatics, and neurobiology. It is a division of Cold Spring Harbor Laboratory, an innovator in life science research and the education of scientists, students, and the public. For more information, visit http://www.cshlpress.com/.

Source: Maria Smit
Cold Spring Harbor Laboratory