Cold Spring Harbor Protocols features methods for
visualizing protein dynamics
Development of Mammalian Cell Lines with lac Operator-Tagged
Chromosomes.
COLD SPRING HARBOR, N.Y. (Wed., Jan. 2, 2008) � 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 (see below).
The first protocoldescribes 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. 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.
Further Information
and Source:
-
Open Access Article:
Yuri G. Strukov and Andrew S. Belmont: Development of Mammalian Cell Lines with lac Operator-Tagged
Chromosomes.
In: CSH Protocols;
2008; doi: 10.1101/pdb.prot4903.
Cold Spring
Harbor Protocols 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.
-
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.
