pegfp n1 clontech pdf

Clontech Laboratories, Inc. A Takara Bio Company 1290 Terra Bella Ave. Mountain View, CA 94043 Technical Support (US) E-mail: [email protected] www.clontech.com Vector Information pEGFP-N3 Vector InformationGenBank Accession #: U57609 Catalog #6080-1 PT3054-5 (PR29967; published 03 October 2002) Description:

pegfp n1 clontech pdf

Clontech Laboratories, Inc. A Takara Bio Company 1290 Terra Bella Ave. Mountain View, CA 94043 Technical Support (US) E-mail: [email protected] www.clontech.com Vector Information pEGFP-1 Vector InformationGenBank Accession #: U55761 Catalog #6086-1 PT3026-5 (PR29973; published 03 October 2002) Restriction Map and Multiple Cloning Site (MCS) of ... 翻訳 · Restriction Map and Multiple Cloning Site (MCS) of pEGFP-N1 Vector. (Unique restriction sites are in bold.) The Not I site follows the EGFP stop codon. The Xba I site (*) is methylated in the DNA provided by CLONTECH. If you wish to digest the vector with this enzyme, you will need to transform the vector into a dam– and make fresh DNA. pEGFP-N1 vector from CLONTECH 4neo-G-CaMP1.6 SV40 polyA CMV IE promoter AflII Kanr/Neor Kozak NheI Bgl II/BamHI(destroied) NotI 4neo-G-CaMP1.6 pN1-4neo-G-CaMP1.6 vector: pEGFP-N1 (CLONTECH: GenGank Accession# U55762) Cutting out: Nhe - NotI Host: DH5α, HB101and other general purpose strains. HindIII and cloned into the pEGFP-N1 vector (Clontech, Mountain View, CA, USA). For pFBL-LacZ-EGFP plasmid construction, the region encoding human FBL was amplified from cDNA (Table S1). The PCR product was digested by BglII and SalI, gel purified and cloned into the pEGFP-N1 vector (Clontech, Mountain View, CA, USA). pEGFP-N1 vector from CLONTECH G-CaMP2 SV40 polyA CMV IE promoter AflII Kanr/Neor Kozak Bgl II NotI G-CaMP2 pN1-G-CaMP2 (pN1-RSET-mG1.6#X-1) vector: pEGFP-N1 (CLONTECH: GenGank Accession# U55762) Cutting out: BglII-NotI Host: DH5α, HB101and other general purpose strains. Selection marker: Kanamycin (30 µg/ml) to E. coli host; Neomycin to ... Molecular Biology of the Cell Vol. 19, 3180–3191, August 2008 The PCH Family Member Proline-Serine-Threonine Phosphatase–interacting Protein 1 Targets to the Leukocyte CELL STRUCTURE AND FUNCTION 42: 131–140 (2017) The Position of the GFP Tag on Actin Affects the Filament Formation in Mammalian Cells Akira Nagasaki 1*, Saku T. Kijima1,2, Tenji Yumoto 3, Miku Imaizumi4, Ayana Yamagishi 1,4, Hyonchol Kim 1,4, Chikashi Nakamura , and Taro Q.P. Uyeda1,2,3 1Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 翻訳 · Additional vectors used to attempt to derive stable GFP expressing cells included pCX-EGFP (supplied by B. Engleward, Massachusetts Institute of Technology) and pEGFP-N1 (Clontech Laboratories, Palo Alto, Calif, USA). pCX-EGFP regulates EGFP (enhanced GFP) under a chicken beta-actin promoter/CMV-IE enhancer and pEGFP-N1 is a sequence variant of pEGFP-N3. Cellular/Molecular Contactin Associates with Sodium Channel Na v1.3 in Native Tissues and Increases Channel Density at the Cell Surface Bhaval S. Shah,1,2,3* Anthony M. Rush,1,2,3* Shujun Liu,1,2,3 Lynda Tyrrell,1,2,3 Joel A. Black,1,2,3 Sulayman D. Dib-Hajj,1,2,3 and Stephen G. Waxman1,2,3 1Department of Neurology and 2The Center for Neuroscience and Regeneration Research, Yale University ... into the multiple cloning site of the pEGFP-N1 plasmid (Clontech) under the control of the cytomegalovirus (CMV) promoter. The recombinant pEGFP-Hsp90a vector was successfully constructed, as confirmed by DNA sequencing analysis. Plaque Assay Plaque assay was used to determine the virus titers [15]. Briefly, 2 Journal of Biomedicine and Biotechnology of an irreversible inhibitor of methyl transferase, C3A human hepatoblastoma cells transfected with GFP showed a signif- (pEGFP-N1, Clontech) was added to each cell suspension at a concentration of 20 μg/ml. Aliquots of 200 μl were then transferred into each of the wells of a 96-well electroporation plate and electroporated with the Gene Pulser MXcell system (Bio-Rad Laboratories, Inc.). subcloned into the pEGFP-N1 vector (Clontech, Takara Bio Inc., Japan) and was named PRDX4t-EGFP. HEK293T cells were ... Activated5 0FlankingRegionofNANOGP8ina Self-RenewalEnvironmentisAssociatedwith IncreasedSphereFormationandTumor GrowthofProstateCancerCells Kai Zhang,1 Marcie Fowler ... 翻訳 · Among the benzodiazepines, midazolam, water-soluble benzodiazepine,6is the most widely used anxiolytic and sedative drug for short procedures and in intensive care. Midazolam is known as a mixed-type agonist of benzodiazepine receptors. In addition to the central neuroinhibitory action of midazolam, it also interferes the synthesis of nitric oxide and tumor necrosis factor-α (TNF-α ... Regulation of miRNA expression during neural cell specification Lena Smirnova,1 Anja Gra¨fe,1 Andrea Seiler,2 Stefan Schumacher,1 Robert Nitsch1 and F. Gregory Wulczyn1 1Center for Anatomy, Institute of Cell Biology and Neurobiology, Charite´ University Hospital, Schumannstraße 20–21, 10098 Berlin, Germany 2National Center for … The pEGFP-N1 construct (Clontech) consists of the cytomegalovirus (CMV) promoter and enhanced GFP coding region. The structures of the fusion genes OlMA1-GFP and OlMA2-GFP, which consist of the 5’ upstream regions of the medaka muscle actin genes and the GFP coding region, were described by Kusakabe et al. (’99; Fig. 1A). constructed by inserting an EGFP fragment of pEGFP-N1 (Clontech) into the EcoRI site of pCAGGS (31). In ovo electroporation Fertile chick eggs obtained from a local farm were incubated at 37°C for 2 days. The eggs were windowed, and 0.1–0.5 mlof PBS containing pCAGGS-EGFP (0.1 mg/ml) and pCAGGS-DsRed (0.1 mg/ml) and siRNA (5 mg/ml) along ... 翻訳 · Immunogenetics (2002) 54:30–38 DOI 10.1007/s00251-001-0416-6 O R I G I N A L PA P E R Aruna P.N. Ambagala · Zhengyu Feng Raul G. Barletta · Subramaniam Srikumaran was inserted at the HindIII and AgeI sites of a pEGFP-Hyg-N1 vector, as previously described (13), which was generated by modifying the commercially available pDsRed-Monomer-Hyg-N1 vector (BD Biosciences Clontech, Mountain View, CA, USA). These plasmids were designed to express EGFP fused to the cytoplasmic actin (Gb′-act) gene, the eGFP gene (derived from pEGFP-N1; Clontech Laboratories Inc., Mountain View, USA) and 3′ untranslated region (UTR), which contained the SV40 polyadenylation signal, were PCR amplified from the plasmid pGact-eGFP (Zhang et al., 2002). tion efficiency was evaluated by means of the pEGFP-N1 plasmid (Clontech Laboratories Inc.), transfected independently, and evaluating the originated green-fluorescent cells. The stable cell lines were isolated, by sub-culturing and selecting with 1 µg/ml of puromycin for 1 month, prior of performing the experiments. 2.5. The pEGFP-N1 construct (Clontech) consists of the cytomegalovirus (CMV) promoter and en-hanced GFP coding region. The structures of the fusion genes OlMA1-GFP and OlMA2-GFP, which consist of the 5’ upstream regions of the medaka muscle actin genes and the GFP coding region, were described by Kusakabe et al. (’99; Fig. 1A). A model of N-terminal Cyclin T1 based on FRETexperiments SERGIO PANTANO†k#, ALESSANDRO MARCELLO‡{#, ARIANNA SABO`§, ALDO FERRARI{, VITTORIO PELLEGRINI{, Journal of Molecular Endocrinology DOI: 10.1530/JME-17-0063 http://jme.endocrinology-journals.org 2017 Society for Endocrinology Printed in Great Britain 翻訳 · Therefore, to determine transfection-induced cell toxicity, the mammalian expression vector pEGFP-N1 (CLONTECH Lab., Palo Alto, CA, USA) has been modified to the dual-cassette expression vectors named pEGFP-Ks by the relocation of its EGFP expression cassette. 翻訳 · Amplify plasmid pEGFP-N1 (Clontech Laboratories Inc., Mountain View, CA, USA) encoding green fluorescent protein (GFP) in DH5α strain of Escherichia coli and isolate it with HiSpeed Plasmid Maxi Kit (Qiagen, Hilden, Germany). cDNA into the EcoRI /BamHI sites of the pEGFP-N1 vector and the SalI/BamHI sites of the pEGFP-C1 vec-tor (CLONTECH Laboratories Inc., Palo Alto, Califor-nia, USA), as described previously (14). Construction of a recombinant adenoviral vector carrying the cPLA 2 cDNA. The human cPLA 2 cDNA was sub-cloned between the Not 1 and Xho1 sites of the ... purification. The 4.7 kb pEGFP-N1 vector (Clontech) was propagated in Escherichia coli and purified using a QIAGEN tip and the associated QIAGEN protocol. Fluo-* To whom correspondence should be addressed. Phone: (219) 631 8632. Fax: (219) 631 6652. E-mail: [email protected] 1 Abbreviations: ANTS, 1-aminonaphthalene-3,6,8-trisulfo- The membrane topological analysis of 3b-hydroxysteroid-D24 reductase (DHCR24) on endoplasmic reticulum Xiuli Lu, Yang Li, Jianli Liu, Xiangyu Cao, Xude Wang, Delong Wang, Hisao Seo1 and Bing Gao2 Department of Biochemistry and Cell Biology, School of Life Science, Liaoning University, Huang-Gu-Qu, Chong-Shang-Zhong-Lu No. 66, Shenyang 110036, China frame with a DNA fragment encoding EGFP, derived from the pEGFP-N1 vector (Clontech Laboratories), in the pCI-bsr vector. PCR was carried out using KOD FX neo DNA polymerase (Toyobo, Osaka, Japan) and primers listed in Supplemental Table 1, and the nucleotide sequences of all PCR-cloned fragments were confirmed by sequencing analysis. chain (eTeNT) with the EGFP derived from pEGFP-N1 vector (Clontech, Mountain View, CA, USA) and the PEST sequence of ornithine decarboxylase, as reported previously (Kinoshita et al., 2012). The transfer plasmid pLV-TRE-EGFP.eTeNT.PEST was based on pFUGW (a gift from D. Baltimore, California Institute Institute). pEGFP-N1 was purchased from Clontech and was used as a control. Transfections were performed by using the Fugene 6 transfection reagent from Roche as specified by the manufacturer. Briefly, Vero cells were grown on glass coverslips in 24-well tissue culture plates, in the absence of antibiotics, until 80% BamHI site of pEGFP-N1 (Clontech). pCHMP6G2A-GFP, which has a point mutation at amino acid 2 (a substitution of alanine for glycine), was obtained by PCR-based site-directed mutagenesis using a QuikChange site-directed mutagenesis kit (Stratagene) accordingtothemanufacturer’sinstructions,andthemutationwas confirmed by DNA sequencing. Brain-derived neurotrophic factor regulates cell motility in human colon cancer Ssu-Ming Huang1,2,3, Chingju Lin4, Hsiao-Yun Lin5, Chien-Ming Chiu2, Chia-Wei Fang2, Kuan-Fu Liao3,6,7, Dar-Ren Chen8 and Wei-Lan Yeh9 1Department of Community Medicine, Preventive Medicine Center, and 2Division of Colon and Rectal Surgery, Department of Surgery, Taichung Tzu Chi Hospital, Buddhist Tzu Chi Medical ... FSP1.GFP mouse was assembled using pEGFP-N1 (Clontech, Palo Alto, California, USA) as a convenience vector. After deletion of the CMV promoter by diges-tion with AgeI and PstI and inserting a linker with XhoI and SfiI restriction sites, the vector was digested with EcoRI (5′) and BamHI (3 ′) in the region 5 ′of the green CLINICAL STUDY Identification and functional characterization of a missense mutation in resistin in two patients with severe obesity and insulin resistance interest, EGFP cDNA (derived from pEGFP-N1; Clontech) The EGFP gene is cloned between the subgenomic promoter and a poly(T) sequence. An alternative Sindbis expression vector, pSinRep(nsp2S726), can be used for prolonged expression with decreased cytotoxicity in dissociatedcultured neurons (see Steps 12 and 13). RESEARCH ARTICLE NanoLuc Reporter for Dual Luciferase Imaging in Living Animals Amanda C. Stacer, Shyam Nyati, Pranav Moudgil, Rahul Iyengar, Kathryn E. Luker, Alnawaz Rehemtulla, and