Access counter started in 7/17/1997
♦ It is great sorrow to announce of Dr. Kaoru Fukami-Kobayashi. (2017.08.05 T.M.)
♦ The IMPC: A global research infrastructure for understanding the role of genes in human development and disease - The 145th RIKEN BRC SEMINAR (2017.08.05 T.M.)
Dr. Terry Meehan
Project co-Leader, Mouse Informatics
European Molecular Biology Laboratory- European Bioinformatics Institute (EMBL-EBI)
Date: Aug 30(Wed.) 2017. 16:00 - (JST)
Place: Moriwaki Hall (BioResource Center Main Building 1F), Tsukuba, Japan
♦ Announcement of bioresources developed by the CRISPR/Cas9 technology (2017.08.01 T.M.)
♦ A new BRC team, Drug-Discovery Cellular Basis Development Team has begun its R&D in Kyoto (2017.07.25 T.M.)
♦ A novel gene involved in germ cell formation commonly discovered in fruit fly and mice (Japanese text only) (2017.02.01 T.M.)
♦ A genetic contamination test for genetically-modified mice at BRC has become available as a simple kit (Japanese text only) (2017.02.01 T.M.)
Distribution of Host E. coli strains for incorporation of synthetic amino acids. (2017/07/25 N.N.)
- The host E. coli strains, RFzero-iy strain and B-95.deltaA strain that can produce recombinant proteins incorporating synthetic amino acids by reassignment of the UAG codon.[more...]
- The RFzero-iy strains are designated to incorporate 3-iodotyrosine into produced proteins. For the incorporation, the strain is transformed with the expression plasmid in which a codon for the target tyrosine is replaced with UAG codon and cultured in 3-iodotyrosine containing medium (Mukai, T. et al., 2011). Two strains based on the BW25113 and BL-21(DE3) are available.
- The B-95.deltaA strains can be assigned their UAG codon to synthetic amino acids by the introduction of plasmids carrying specific pair of UAG-reading tRNA and an aminoacyl-tRNA synthetase (aaRS) variant*. Increased productivity of sulfonated hirudin which is expected to improve the inhibition of blood coagulation was reported (Mukai, T. et al., 2015). The original and the growth improved derivative strains are available.
Mukai, T. et al., Biochem. Biophys. Res. Commun. 411 (4): 757-761, 2011. [PMID 21782790]
Mukai, T. et al., Sci. Rep. 5: 9699, 2015. [PMID 25982672]
Incorporation of synthetic amino acids into proteins at specific sites
- DNA Resource
BW25113-based RFzero-iy (cat # RDB14427)
BL21(DE3)-based RFzero-iy (cat # RDB14428)
B-95.deltaA (cat # RDB13711)
B-95.deltaAdeltafabR (cat # RDB13712)
*Plamids that introduce synthetic amino acids are provided by RIKEN CLST.
Evaluation of chaperone mediated autophagy (CMA) activity by the GAPDH-HT indicator. (2017/06/29 N.N.)
- "GAPDH-HT indicator" is a CMA marker consisting of HaloTag and GAPDH which is a typical substrate of CMA. The CMA activity in the cells can be evaluated by using GAPDH-HT indicator and an appropriate under the fluorescent-labeled HaloTag ligand.[more]
Seki, T. et al. Establishment of a novel fluorescence-based method to evaluate chaperone-mediated autophagy in a single neuron. PLoS One 7 (2): e31232, 2012. PMID: 22363588
Sato, M. et al. Fluorescent-based evaluation of chaperone-mediated autophagy and microautophagy activities in cultured cells. Genes Cells 21 (8): 861-873, 2016. PMID: 27377049
- DNA Resource
GAPDH-HT/pcDNA5/FRT (cat # RDB15088)
Establishment of deficient cells using Cre recombinase and floxed mouse derived cells (2017/06/23 T.M.)
- Has2 deficient cells were established by using Cre recombinase expressing AxCANCre adenovirus. Mouse Has2 gene floxed allele cells were segregated from the transgenic mice having the exon 2 flanked by two loxP sites. And then the cells were infected with AxCANCre adenovirus carrying the Cre recombinase to remove the exon 2.[more]
Chanmee, T. et al. Hyaluronan production regulates metabolic and cancer stem-like properties of breast cancer cells via hexosamine biosynthetic pathway-coupled HIF-1 signaling. J. Biol. Chem. 291 (46): 24105-24120, 2016. PMID: 27758869.
- DNA Resource
AxCANCre (cat# RDB01748)
M-INK, a novel tool for tagging the mature melanosomes.(2017/05/02 N.N.)
- A novel tool "M-INK" to visualize melanosome in melanocyte and keratinocyte has arrived.[more]
G-CaMP, a fluorescent calcium sensor.(2017/03/16 N.N.)
- A Fluorescent probes "G-CaMP" for measuring intracellular calcium concentration are available.[more]
- The G-CaMPs are calcium sensor proteins consisting of Calmodulin (CaM) calcium-binding region, M13 fragment of myosin light chain kinase and fluorescent protein. In the original paper, G-CaMPs were used to observe cardiomyocytes differentiated from iPS cells (Shiba, Y. et al, 2016) and neurons of zebrafish (Muto, A. et al, 2011).
- Shiba, Y. et al. Allogeneic transplantation of iPS cell-derived cardiomyocytes regenerates primate hearts. Nature, 538 (7625): 388-391, 2016. PMID: 27723741
- Ohkura, M. et al. Genetically encoded green fluorescent Ca2+ indicators with improved detectability for neuronal Ca2+ signals. PLoS One, 7 (12): e51286, 2012. PMID: 23240011
- Ohkura, M. et al. An improved genetically encoded red fluorescent Ca2+ indicator for detecting optically evoked action potentials. PLoS One, 7 (7): e39933, 2012. PMID: 22808076
- Muto, A. et al. Genetic visualization with an improved GCaMP calcium indicator reveals spatiotemporal activation of the spinal motor neurons in zebrafish. Proc. Natl. Acad. Sci. U S A., 108 (13): 5425-5430, 2011. PMID: 21383146
- DNA Resource
G-CaMP4.1 (cat.# RDB14606)
G-CaMP-HS (cat.# RDB14607)
G-CaMP6 (cat.# RDB14609)
G-CaMP7 (cat.# RDB14610)
G-CaMP8 (cat.# RDB14611)
G-CaMP7.09 (cat.# RDB14612)
Construction of reporter gene having long promoter by using BAC clone and with recombineering technique.(2017/02/28 T.M.)
- Luciferase reporter assay is a powerful tool for real-time monitoring of gene expression in living cells. It is recommended to construct reporter gene with lager promoter but hard to do so by means of common cloning technique. In this article, 20 kb promoter of Hprt gene was retrieved and cloned into a luciferase reporter by using BAC clone and with recombineering technique.[more]
A fluorescent probe to evaluate activity of autophagy. (2017/02/21 N.N.)
- Plasmid clone of a novel probe capable of evaluating the activity of autophagy by GFP/RFP signal ratio has arrived.[more]
Green- and Red-emitting luciferases with luminescence. (2017/2/2 T.M.)
- Now, we offer five novel luciferase genes of fireflies. The luciferases hold high luminescence and stability in a wide pH range.
- Deposited Resource
♦ Histac-H3K9/K14 for visualization of acetylation activity of histone H3 in cells.(2016/9/27 T.M.)
Nakaoka, S., Sasaki, K., Ito, A., Nakao, Y., Yoshida, M. A Genetically Encoded FRET Probe to Detect Intranucleosomal Histone H3K9 or H3K14 Acetylation Using BRD4, a BET Family Member. ACS Chem. Biol. 11 (3): 729-733, 2016. PMID: 25946208
♦ GimRET for visualization of protein concentration in cells.(2016/9/13 T.M.)
Morikawa, J.T., Fujita, H., Kitamura, A., Horio, T., Yamamoto, J., Kinjo, M., Sasaki, A., Machiyama, H., Yoshizawa, K., Ichimura, T., Imada, K., Nagai, T., Watanabe, M.T. Dependence of fluorescent protein brightness on protein concentration in solution and enhancement of it. Sci. Rep. 6: 22342, 2016. PMID: 26956628
♦ Nakanori that specifically binds to a complex of sphingomyelin and cholesterol.(2016/9/13 T.M.)
Makino, A. et al. A novel sphingomyelin/cholesterol domain-specific probe reveals the dynamics of the membrane domains during virus release and in Niemann-Pick type C. FASEB. J., 2016. Aug, 4. In press. PMID: 27492925.
♦ Fluorescent Ubiquitination-based Cell Cycle Indicator (Fucci) by single plasmid.(2016/6/20 T.M.)
- Fucci2a cell cycle phase markers allow visualization and estimation of cell cycle progress by observation of green and red fluorescent proteins. [more...]
- This plasmid express two distinct colored fluorescent proteins, one of which fused to Cdt1 degron, another one of which fused to Geminin degron. Therefore, the transfectant indicate distinct color fluorescence between the cell (G1) and the cell (S/G2/ M).
Mort, R.L., Ford, M.J., Sakaue-Sawano, A., Lindstrom, N.O., Casadio, A., Douglas, A.T., Keighren, M.A., Hohenstein, P., Miyawaki, A., Jackson, I.J. Fucci2a: a bicistronic cell cycle reporter that allows Cre mediated tissue specific expression in mice. Cell Cycle 13 (17): 2681-2696, 2014.
- DNA Resource
Transgenic mouse B6;129-Gt(ROSA)26Sor (cat.# RBRC06511) is also available from the Experimental Animal Division.
- pCAG-Fucci2a (cat.# RDB13080) for transient expression of Fucci2a marker.
- pROSA-floxNeo-Fucci2a (cat.# RDB13081) for targeted insertion of Fucci2a marker into mouse ROSA26 locus.
♦ Genius method to get rid of the target protein with human culture cells (2016.03.25 K.N.)
Now available the plasmid for the system of Auxin-indusible degradation (AID) tag of your target protein in human cells from RIKEN BRC. [more...]
Quality Examination and Information Release Policy on Our Bioresources (2014.09.01)
♦ Announcement of the interruption on our mail server (Mar 01, 2017)
♦ Prepayment of distribution fees (overseas organizations only)
♦ New Information from RIKEN BioResource Center
RIKEN and all those who have deposited materials or information are not liable for any harm to any individual that might arise as a result of the use of any resources or informaion in the RIKEN databases anywhere in the world.
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