What is H2B GFP?
Conclusions: The H2B-GFP system allows the high-resolution imaging of chromosomes, including DMs, without compromising nuclear and chromosomal structures and has revealed the distinctive clustering behavior of DMs in mitotic cells which contributes to their asymmetric distribution to daughter cells.
What is a GFP reporter?
GFP from the jellyfish Aequoria victoria is a versatile reporter for monitoring gene expression in vivo, in situ and in real time. GFP fluoresces bright-green upon exposure to UV light, unlike other bioluminescent reporters which require additional proteins, substrates or cofactors to emit light.
What is a GFP assay?
Green fluorescent protein (GFP) is a powerful tool for cell-based assays owing to the intrinsic fluorescence of this protein that allows real-time analysis of molecular events in living cells.
What is H2B mCherry?
Maize histone H2B-mCherry: a new fluorescent chromatin marker for somatic and meiotic chromosome research. DNA Cell Biol.
Why GFP is a popular reporter gene?
Two advantages of GFP as a reporter of gene expression are that i. protein accumulation can be directly observed in living cells prior to quantitative analysis, and ii. GFP gene expression can be measured on a cell-by-cell basis.
What is a reporter gene used for?
Reporter genes are genes whose products can be readily assayed subsequent to transfection, and can be used as markers for screening successfully transfected cells, for studying regulation of gene expression, or serve as controls for standardizing transfection efficiencies.
How do you test for green fluorescent proteins?
The GFP expression may be detected by fluorescence microscopy, fluorescence-activated cell sorting (FACS) analysis, or fluorometer assays 24–72 h posttransfection, depending on the host cell line used. There is one published report of a stable mammalian cell line expressing GFP (48).
What is H2B GFP fusion protein used for?
The H2B–GFP fusion protein becomes incorporated into nucleosomes without affecting cell cycle progression [7]. The H2B-GFP fusion protein makes the fluorescent label very stable and the nuclear localization also facilitates easier identification of label-retaining cells in histological sections.
What is H2B-GFP fluorescence?
When the H2B-GFP mice are crossed to mice harboring a promoter-driven tetracycline-regulatable transactivator protein, H2B-GFP fluorescence is produced when the tTA binds to the TRE. The H2B–GFP fusion protein becomes incorporated into nucleosomes without affecting cell cycle progression.
Can H2B-GFP mice be used to fluorescently label HSCs in vivo?
Because there is no one marker that is uniquely and specifically expressed by HSCs, we sought to use the H2B-GFP mice to fluorescently label HSCs in vivo.
What is H2B-GFP mice?
The group that generated the H2B-GFP mice used a keratin 5 promoter-driven tetracycline-regulated transactivator to specifically label the skin. After a 4-week doxycycline chase, the only GFP+cells were in the bulge, the presumptive skin stem cell compartment.