The goal of the imsr is to assist the international scientific community in locating and obtaining mouse resources for research. Gfp fused gene pegfpc2 not detected by gfp mouse antibodies in western blotting any ideas. Mgisoftware developer tools for the mouse genome informatics. Morphological evidence of autophagy in human neurological diseases 160 1. Transgenic mice with green fluorescent proteinlabeled pancreatic. Adult mouse pomcgfp mhypoapomcgfp14 and mpitapomcgfp1 the mhypoapomcgfp cell lines were generated using a proprietary platform technology enabling the creation of pools of immortalized hypothalamic neurons.
Egfp is a basic constitutively fluorescent green fluorescent protein published in 1996, derived from aequorea victoria. Steps to create prebuilt human hg19, grch38, mouse mm10. The jackson laboratory handbook on genetically standardized mice. Jun 25, 2019 brca2 is a key homologous recombination mediator in vertebrates. They also find that hsf2bp contributes to dna repair in mouse embryonic stem cells. Green fluorescent protein gfp thermo fisher scientific.
Green fluorescent protein gfp is a novel genetic reporter system derived from the bioluminescent jellyfish aequorea victoria which, when expressed in either eukaryotic or prokaryotic cells and illuminated by blue or uv light, yields a bright green fluorescence chalfie et al. Government furnished property includes, but is not limited to, spares and property furnished for repairs, maintenance, overhaul, or modification. A recent example of a knockin transgenic mouse is the mirr144451 gfp line, in which gfp is expressed in adult and fetal liver erythroid cells 26, 27 figure 3. Gfp 4b10 mouse mab detects gfp, yfp, and cfptagged proteins exogenously expressed in cells. Fulllength blots are presented in supplementary fig. This implies that i will probably get gfp homozygous mice and i have heard that this can carry. This transgenic mouse line, with an enhanced gfp egfp cdna under the control of a chicken betaactin promoter and cytomegalovirus enhancer, have widespread egfp fluorescence, with the exception of erythrocytes and hair. The deleted region encoded an aminoterminal portion of the protein that is crucial for. Joseph gleesons lab contains the insert foxg1 and is published in nat med. Softmouse is the leading brand in online mouse colony management webbased and mobile solutions. Generation of transgenic mouse fluorescent reporter lines. National institute of neurological disorders and stroke, contract. In vivo imaging of cx3cr1gfpgfp reporter mice with spectral.
Tiempo realtime acquisition software olympus was used to collect. Jan 30, 2020 in mouse, we carried out bulk rnaseq of macrophages isolated from p1, p7 and adult hearts by facs at baseline and days 1, 4 and 7 postmi supplementary fig. As storytellers who rely on images or video to report on. Mgi is the international database resource for the laboratory mouse, providing integrated genetic, genomic, and biological data to facilitate the study of human health and disease. Welcome to the yeast gfp fusion localization database the yeastgfp database of global analysis of protein localization studies in the budding yeast, s. Plasmid mouse ab crystallin 800bp promoteracgfp from dr. For example, green fluorescent es cells should increase the efficiency of chimera production by allowing the preselection of chimeric blastocysts prior to transfer into. Crossing gad1 tm1tama with mice expressing cre recombinase in the germline removed the loxpflanked neo.
Functional dissection of hematopoietic stem cell populations. Database search to search the database, enter the text in the box below and choose the field to search on. Our mission is to discover precise genomic solutions for disease and empower the global biomedical community in the shared quest to improve human health. Zfin provides a wide array of expertly curated, organized and crossreferenced zebrafish research data. Western blot demonstrated that the cerebral cortex contained more gfp than the cerebral cortex of animals with the neo present. In order to establish sufficient progress is being. Alpha yaps lab contains the insert ecadherin and is published in j cell sci. Sep 12, 2017 hematopoietic stem cells hscs in a steady state can be efficiently purified by selecting for a combination of several cell surface markers.
Green fluorescent protein gfp is a versatile biological marker for monitoring physiological processes, visualizing protein localization, and detecting transgenic expression in vivo. The cd44egfp mice should provide a valuable resource for electrophysiological and connectivity studies of amacrine cells in the mouse retina. Over 2 million visitors each month use driverguide to solve driver problems. A portal to genespecific content based on ncbis refseq project, information from model organism databases, and links to other resources. Gfp and rfpexpressing brain tumors implanted in the brain in a. Mouse anticgfptag monoclonal antibody can be used to detect cgfp fusi. Spectral domain optical coherence tomography sdoct and scanning laser ophthalmoscopy slo are extensively used in experimental ophthalmology. Use with snapgene software or the free viewer to visualize additional data and align other sequences. I have just started to generate a gfp mice colony and i was wondering if it is okay to cross two. Cf dyes offer exceptional brightness and photostability. Mmt gfp rfp cells were inject ed in the footpad of gfp transgenic nude mice. Can anyone recommend a mouse colony management software. B16f0 gfp mouse melanoma cells were injected into the tail vein or. An inventory of mammalian mitochondrial genes broad.
Gfp has been tweaked for codon usage, expression, and fluorescence. Take advantage of our large inventories of common inbred strains and the convenience of having your breeding and drug efficacy needs met by the leading experts in mouse modeling. A software tool for the quantification of metastatic colony growth. Although a bright green light emission was observed in the muscle and pancreas of more than 20 of these transgenic mouse lines, gfp expression was not ubiquitous and light emission was not. Transgenic mice expressing fluorescent proteins at various points in the hematopoietic hierarchy, from hematopoietic stem cell to multipotent progenitors to each of the final differentiated cell types, have provided valuable tools for tagging, tracking, and isolating these cells. Mason posners lab contains the insert alpha bcrystallin promoter 800 bp fragment, mus musculus and is published in peerj. The jackson laboratory is an independent, nonprofit organization focusing on mammalian genetics research to advance human health. Simply the best webbased mouse colony database software and mobile laboratory information management system lims in the cloud. We have established new human and rodent tumors that stably express very high levels of the aequorea victoria green fluorescent protein gfp and transplanted these to appropriate animals. A concern for the knockin approach is that it may result in haploinsufficiency, influencing not only expression of the fp reporter but also the phenotype of the resulting animal.
The plant organellome database a database of transgenic plants or cultured cells, mostly arabidopsis, expressing fluorescent proteintagged proteins to visualize organellar locations. A, wholebody image of untrea ted mmt gfp rfp cells in the footpad of a live gfp mouse. In the present protocol, mice expressing green fluorescent protein gfp under the promoter of cx3cr1 balbccx3cr1 gfp gfp were used to image microglia cells in vivo in the retina. Analysis of the amino acid sequence of lrrc32 by signalp 3.
Creating a reference package with cellranger mkref software. Green fluorescent protein has been engineered to produce a vast number of variously colored mutants, fusion proteins, and biosensors. The use of a gfp antibody for research applications in transgenic c. A bac clone containing the entire pvalb gene and approximately 150 kb of upstream and 25 kb downstream regions was used for bac modifications.
Gfp monoclonal antibody antigfp monoclonal a ntibody is provided at a concentration of 1 mg ml recognizes green fluorescent protein and its variants including egfp, yfp, eyfp and cfp. Lk3 transgenic mice mus musculus linnaeus, 1758 mus sp. Try softmouse forever free mouse colony management software or premium. Does anyone have a vglut2gfp mouse with good cell body expression in the. The gfp expressing cells in the gcl resemble the type b1, or possibly a2 ganglion cells.
Mgimouse genome informaticsthe international database. Bd facschorus tm intuitively designed software simplifies sorting by using a streamlined workflow that eliminates manual setup and monitoring, so that you can focus on the science. The coral green fluorescent protein cgfp from vector prnatinh1. C controls for pla included i cells were incubated with alpha gfp anti mouse and alpha gfp antirabbit to serve as a positive control. Hsf2bp interacts with a conserved domain of brca2 and is. A gfp expression cassette was inserted in the first coding exon at the translation initiation site. Hela cell transfected with pshooter pcmvmycmito gfp, then fixed and permeabilized.
These transgenic mice express enhanced green fluorescent protein gfp under the direction of the human ubiqutin c promoter in all tissues examined. Read here what the gfp file is, and what application you need to open or convert it. Does anyone have experience in visualizing egfp mice. The zebrafish information network zfin is the database of genetic and genomic data for the zebrafish danio rerio as a model organism. This product is prepared by labeling high quality monoclonal mouse antigfp with a selection of fluorescent cf dyes. This transgenic mouse line, with an enhanced gfp egfp cdna under the control of a chicken betaactin promoter and cytomegalovirus enhancer, have widespread egfp fluorescence, with the exception of. Chapter 8 using genetic mouse models to study the biology. The bestlabeled gfp expressing cell type in the inl was a widefield amacrine cell that ramified in stratum 3. Generation of transgenic mouse fluorescent reporter lines for. These mice can be used as a source of marker cells that are fluorescent green. During the development of the hematopoietic system, at least 8 distinct lineages are generated in the mouse embryo. The jellyfish aequorea victoria contains green fluorescent protein gfp tag. This mutant mouse strain represents a useful tool in studies related to hematopoietic cell.
A webbased lab mouse colony laboratory information management software system. Five transgenic founders were identified by pcr and confirmed by southern blotting. Green fluorescent protein is a 27 kda protein produced from the jellyfish aequorea victoria, which emits green light emission peak at a wavelength of 509nm when excited by blue light. Wholebody, noninvasive, subcellular imaging of drug response of dualcolor mouse mammary cancer cells and gfp stromal cells. Input a list of ids or gene symbols and retrieve other database ids and gene attributes e. Fred gages lab contains the insert hb9 gfp and is published in cell stem cell. It was soon after established that the emission spectra of gfp was right around 509nm, or. Currently five proteomes are available including human, mouse, c.
I would, as proposed before, check that both your transduced astrcocytes and your mouse brains express gfp by qpcr or just pcr, to be sure that the construct. The imsr is a searchable online database of mouse strains, stocks, and mutant es cell lines available worldwide, including inbred, mutant, and genetically engineered strains. The endogenous locus was disrupted by the insertion of sequence encoding green fluourescent protein gfp, replacing the first 390 bp of the coding exon exon 2. Can anyone recommend a mouse colony management software system. Hematopoietic stem cells hscs in a steady state can be efficiently purified by selecting for a combination of several cell surface markers. Anti gfp and betaactin antibodies were used to detect gfp and betaactin, respectively. This resource is powered by the protein data bank archiveinformation about the 3d shapes of proteins, nucleic acids, and complex assemblies that helps students and researchers understand all aspects of biomedicine and agriculture, from protein synthesis to health and disease. Optimized codon usage and chromophore mutations provide enhanced sensitivity with the green fluorescent protein most of the mammalian expression vectors would use egfp eenhanced rather than the original jellyfish version. Tetracyclinecontrolled transgene activation using the rosa26. In vivo imaging of cx3cr1gfpgfp reporter mice with.
Scientific research is complex, but sorting doesnt have to be. Wholebody optical imaging of green fluorescent protein. We therefore modified our expression construct to include the enhanced green fluorescent protein egfp. Please note that the gfp, yfp, and cfp tags add approximately 27 kda to the molecular weight of the fusion protein. As a member of the wwpdb, the rcsb pdb curates and annotates pdb data. The neo cassette was removed through subsequent flpmediated recombination. Gfp can be excited by the 488 nm laser line and is optimally detected at 510 nm. Build a custom reference with cellranger mkref software. Gensat brain atlas of gene expression in egfp transgenic mice.
Basic steps in the generation of a fluorescent transgenic reporter mouse line. I currently use the software to manage a colony of around 300 mice, including 43 breeding pairs and without it ill be completely lost. How to use gfptagged proteins in cell biology namboori b. Monoclonal antigfp is designed to detect enhanced gfp and gfptagged recombinant proteins. Intravital frap imaging using an ecadheringfp mouse. Pdf transgenic mice with green fluorescent proteinlabeled. The linker database was developed by richard george. Download drivers for free from the webs largest driver. Plasmid sequence and snapgene enhanced annotations. Gfp is defined as property in the possession of, or directly acquired by, the government and subsequently furnished to the contractor for performance of a contract.
To this end, the ecadherin gfp mouse could also be used to decipher other molecular pathways downstream of mutant p53 to be targeted in future studies or be used with other kras or p53 mutations to determine whether ecadherin mobilization is a general phenomenon for all mutant p53 variants in this disease muller et al. Using genetic mouse models to study the biology and pathology of autophagy in the central nervous system zhenyu yue, gay r. Softmouse mouse colony management software online database and mouse breeding software in the cloud. Mouse anti gfp antibody is functional by western blot, elisa, immunofluorescene micorscopy and immunohistochemistry. Since 1996, experts recommend driverguide to fix pc problems fast. Pdf transgenic mice with green fluorescent protein. The most efficient lims system to manage your mouse colony data. The green fluorescent protein gfp is responsible for the green bioluminescence of. Beside the above main applications we envision other important uses for our gfp tagged escelltransgenic mouse system.
Generating green fluorescent mice by germline transmission of. Characterization of green fluorescent proteinexpressing. Certain hematopoietic cell types display distinct expression levels of gfp, allowing identification of different cells types by facs analysis. It is reported to be a rapidlymaturing weak dimer with moderate acid sensitivity. Ncbi reference sequence database a comprehensive, integrated, nonredundant, wellannotated set of reference sequences including genomic, transcript, and protein. If you ask a biologist, there should be only one gfp sequence at least in the research lab setting. Download drivers for free from the webs largest driver database. A loxp site flanked gfra1 cdna with sv40 polya signal sequences, a gfp gene with polya signal sequences, and an frt site flanked neomycin resistance gene cassette were introduced into exon 2. Generating green fluorescent mice by germline transmission. Thermo fisher antibodies are validated for applications including western blotting, immunocytochemistry, flow cytometry, and chromatin immunoprecipitation.
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