o***e
发帖数: 130 | 1 我想看一个蛋白在不同cell cycle(G0, G1, S, M)时期的distribution。用
fluorescence microscopy, 有什么marker或其他分辨方法吗？谢谢！
实在不行可以先sort，是下策了。 | m*******r
发帖数: 202 | 2 Cell Cycle. 2005 Mar;4(3):453-5. Epub 2005 Mar 7.
Cell cycle markers for live cell analyses.
Easwaran HP, Leonhardt H, Cardoso MC.
SourceMax Delbrück Center for Molecular Medicine, Berlin, Germany.
Abstract
Many cellular processes are regulated by cell cycle dependent changes in
protein dynamics and localization. Studying these changes in vivo requires
methods to distinguish the different cell cycle stages. Here we demonstrate
the use of DNA Ligase I fused to DsRed1 as an in situ marker to identify S
phase and the subsequent transition to G2 in live cells. Using this marker,
we observed changes in the nuclear distribution of Dnmt1 during cell cycle
progression. Based on the different nuclear distribution of DNA Ligase I and
Dnmt1 in G2 and G1, we demonstrate that the combination of both proteins
allows the direct discrimination of all cell cycle phases using either
immunostainings or fusions with fluorescent proteins. These markers are new
tools to directly study cell cycle dependent processes in both, fixed and
living cells.
Chembiochem. 2010 May 17;11(8):1037-47.
Cell-cycle markers and biosensors.
Kurzawa L, Morris MC.
SourceUniversité de Montpellier, Montpellier, France.
Abstract
Since the first schematic illustrations of dividing cells, we have come a
long way in characterising eukaryotic cells and defining their cell-cycle
status thanks to a number of complementary approaches. Although most of
these approaches rely on cell-fixation procedures to identify molecular
components in cell lysates, cultured cells or tissues, the development of
GFP technology has enabled visualisation of virtually any fusion protein in
cellulo and in vivo, and the exploitation of functional elements with well-
defined spatiotemporal characteristics has enabled the development of
genetically encoded fluorescent markers of cell-cycle phases, thus providing
novel means of characterising the status of living cells in real time with
high resolution. Together with technological advances in fluorescence
chemistry and imaging approaches, the more recent development of fluorescent
biosensors has provided direct means of probing cell-cycle regulators and
of studying their dynamics with high spatial and temporal resolution. Here
we review classical approaches that rely on cell fixation to characterise
the cell-cycle status and its regulatory enzymes, and we describe the more
recent development of cell-cycle markers based on genetically encoded
fusions of fluorescent proteins with characteristic cell-cycle features, and
of fluorescent biosensor technology to probe cell-cycle regulators in
living cells. Biosensors not only provide a means of characterising the
behaviour of cell-cycle regulators in their natural environment, they are
also very useful for comparative studies of biological processes in healthy
and pathological conditions, and can be further applied to diagnostic
approaches to assess the status of a specific target, and to monitor
response to therapeutic intervention.
【在 o***e 的大作中提到】
: 我想看一个蛋白在不同cell cycle(G0, G1, S, M)时期的distribution。用
: fluorescence microscopy, 有什么marker或其他分辨方法吗？谢谢！
: 实在不行可以先sort，是下策了。
|
|
