Simultaneous fluorescence-activated cell sorter analysis of two distinct transcriptional elements within a single cell using engineered green fluorescent proteins.

Abstract:

:Green fluorescent protein (GFP) is widely used as a reporter gene in both prokaryotes and eukaryotes. However, the fluorescence levels of wild-type GFP (wtGFP) are not bright enough for fluorescence-activated cell sorting or flow cytometry. Several GFP variants were generated that are brighter or have altered excitation spectra when expressed in prokaryotic cells. We engineered two GFP genes with different combinations of these mutations, GFP(S65T,V163A) termed GFP-Bex1, and GFP(S202F,T203I,V163A) termed GFP-Vex1. Both show enhanced brightness and improved signal-to-noise ratios when expressed in mammalian cells and appropriately excited, compared with wtGFP. Each mutant retains only one of the two excitation peaks of the wild-type protein. GFP-Bex1 excites at 488 nm (blue) and GFP-Vex1 excites at 406 nm (violet), both of which are available laser lines. Excitation at these wavelengths allows for the independent analyses of these mutants by fluorescence-activated cell sorting, permitting simultaneous, quantitative detection of expression from two different genes within single mammalian cells.

authors

Anderson MT,Tjioe IM,Lorincz MC,Parks DR,Herzenberg LA,Nolan GP,Herzenberg LA

doi

10.1073/pnas.93.16.8508

subject

Has Abstract

pub_date

1996-08-06 00:00:00

pages

8508-11

issue

16

eissn

0027-8424

issn

1091-6490

journal_volume

93

pub_type

杂志文章
  • Algorithmic design of self-assembling structures.

    abstract::We study inverse statistical mechanics: how can one design a potential function so as to produce a specified ground state? In this article, we show that unexpectedly simple potential functions suffice for certain symmetrical configurations, and we apply techniques from coding and information theory to provide mathemat...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0901636106

    authors: Cohn H,Kumar A

    更新日期:2009-06-16 00:00:00

  • Cooperative enhancement of specificity in a lattice of T cell receptors.

    abstract::Two of the most important models to account for the specificity and sensitivity of the T cell receptor (TCR) are the kinetic proofreading and serial ligation models. However, although kinetic proofreading provides a means for individual TCRs to measure accurately the length of time they are engaged and signal appropri...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.101113698

    authors: Chan C,George AJ,Stark J

    更新日期:2001-05-08 00:00:00

  • Immortalized keratinocyte lines derived from human embryonic stem cells.

    abstract::Cells of the human embryonic stem (hES) cell line H9, when cultured in the form of embryoid bodies, give rise to cells with markers of the keratinocyte of stratified squamous epithelia. Keratinocytes also form in nodules produced in scid mice by injected H9 cells; the hES-derived keratinocytes could be recovered in cu...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0510953103

    authors: Iuchi S,Dabelsteen S,Easley K,Rheinwald JG,Green H

    更新日期:2006-02-07 00:00:00

  • Protease nexin-2/amyloid beta-protein precursor limits cerebral thrombosis.

    abstract::The amyloid beta-protein precursor (AbetaPP) is best known as the parent molecule to the amyloid beta-peptide that accumulates in the brains of patients with Alzheimer's disease. Secreted isoforms of AbetaPP that contain the Kunitz proteinase inhibitor domain are analogous to the previously identified cell-secreted pr...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0507798102

    authors: Xu F,Davis J,Miao J,Previti ML,Romanov G,Ziegler K,Van Nostrand WE

    更新日期:2005-12-13 00:00:00

  • Plant-derived anti-Lewis Y mAb exhibits biological activities for efficient immunotherapy against human cancer cells.

    abstract::Although current demands for therapeutic mAbs are growing quickly, production methods to date, including in vitro mammalian tissue culture and transgenic animals, provide only limited quantities at high cost. Several tumor-associated antigens in tumor cells have been identified as targets for therapeutic mAbs. Here we...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0603043103

    authors: Brodzik R,Glogowska M,Bandurska K,Okulicz M,Deka D,Ko K,van der Linden J,Leusen JH,Pogrebnyak N,Golovkin M,Steplewski Z,Koprowski H

    更新日期:2006-06-06 00:00:00

  • Characterization of a murine type II sodium-phosphate cotransporter expressed in mammalian small intestine.

    abstract::An isoform of the mammalian renal type II Na/Pi-cotransporter is described. Homology of this isoform to described mammalian and nonmammalian type II cotransporters is between 57 and 75%. Based on major diversities at the C terminus, the new isoform is designed as type IIb Na/Pi-cotransporter. Na/Pi-cotransport mediate...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.95.24.14564

    authors: Hilfiker H,Hattenhauer O,Traebert M,Forster I,Murer H,Biber J

    更新日期:1998-11-24 00:00:00

  • Recreational fishing selectively captures individuals with the highest fitness potential.

    abstract::Fisheries-induced evolution and its impact on the productivity of exploited fish stocks remains a highly contested research topic in applied fish evolution and fisheries science. Although many quantitative models assume that larger, more fecund fish are preferentially removed by fishing, there is no empirical evidence...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1212536109

    authors: Sutter DA,Suski CD,Philipp DP,Klefoth T,Wahl DH,Kersten P,Cooke SJ,Arlinghaus R

    更新日期:2012-12-18 00:00:00

  • Glucose intolerance caused by a defect in the entero-insular axis: a study in gastric inhibitory polypeptide receptor knockout mice.

    abstract::Mice with a targeted mutation of the gastric inhibitory polypeptide (GIP) receptor gene (GIPR) were generated to determine the role of GIP as a mediator of signals from the gut to pancreatic beta cells. GIPR-/- mice have higher blood glucose levels with impaired initial insulin response after oral glucose load. Althou...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.96.26.14843

    authors: Miyawaki K,Yamada Y,Yano H,Niwa H,Ban N,Ihara Y,Kubota A,Fujimoto S,Kajikawa M,Kuroe A,Tsuda K,Hashimoto H,Yamashita T,Jomori T,Tashiro F,Miyazaki J,Seino Y

    更新日期:1999-12-21 00:00:00

  • Modulation of NKG2D, NKp46, and Ly49C/I facilitates natural killer cell-mediated control of lung cancer.

    abstract::Natural killer (NK) cells play a critical role in controlling malignancies. Susceptibility or resistance to lung cancer, for example, specifically depends on NK cell function. Nevertheless, intrinsic factors that control NK cell-mediated clearance of lung cancer are unknown. Here we report that NK cells exposed to exo...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1804931115

    authors: Shi L,Li K,Guo Y,Banerjee A,Wang Q,Lorenz UM,Parlak M,Sullivan LC,Onyema OO,Arefanian S,Stelow EB,Brautigan DL,Bullock TNJ,Brown MG,Krupnick AS

    更新日期:2018-11-13 00:00:00

  • Unveiling functional protein motions with picosecond x-ray crystallography and molecular dynamics simulations.

    abstract::A joint analysis of all-atom molecular dynamics (MD) calculations and picosecond time-resolved x-ray structures was performed to gain single-molecule insights into mechanisms of protein function. Ensemble-averaged MD simulations of the L29F mutant of myoglobin after ligand dissociation reproduce the direction, amplitu...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0405295101

    authors: Hummer G,Schotte F,Anfinrud PA

    更新日期:2004-10-26 00:00:00

  • Double proton transfer dynamics of model DNA base pairs in the condensed phase.

    abstract::The dynamics of excited-state double proton transfer of model DNA base pairs, 7-azaindole dimers, is reported using femtosecond fluorescence spectroscopy. To elucidate the nature of the transfer in the condensed phase, here we examine variation of solvent polarity and viscosity, solute concentration, and isotopic frac...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0702944104

    authors: Kwon OH,Zewail AH

    更新日期:2007-05-22 00:00:00

  • Optogenetic fMRI interrogation of brain-wide central vestibular pathways.

    abstract::Blood oxygen level-dependent functional MRI (fMRI) constitutes a powerful neuroimaging technology to map brain-wide functions in response to specific sensory or cognitive tasks. However, fMRI mapping of the vestibular system, which is pivotal for our sense of balance, poses significant challenges. Physical constraints...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1812453116

    authors: Leong ATL,Gu Y,Chan YS,Zheng H,Dong CM,Chan RW,Wang X,Liu Y,Tan LH,Wu EX

    更新日期:2019-05-14 00:00:00

  • Genomewide transcriptional changes associated with genetic alterations and nutritional supplementation affecting tryptophan metabolism in Bacillus subtilis.

    abstract::DNA microarrays comprising approximately 95% of the Bacillus subtilis annotated protein coding ORFs were deployed to generate a series of snapshots of genomewide transcriptional changes that occur when cells are grown under various conditions that are expected to increase or decrease transcription of the trp operon se...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1031606100

    authors: Berka RM,Cui X,Yanofsky C

    更新日期:2003-05-13 00:00:00

  • Inhibition of host translation by virus infection in vivo.

    abstract::Infection of cultured cells with lytic animal viruses often results in the selective inhibition of host protein synthesis, whereas viral mRNA is efficiently translated under these circumstances. This phenomenon, known as "shut off," has been well described at the molecular level for some viruses, but there is not yet ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1004110107

    authors: Toribio R,Ventoso I

    更新日期:2010-05-25 00:00:00

  • On the mechanism of action of aldosterone.

    abstract::Studies to elucidate the mode of action of aldosterone have been carried out in the amphibian urinary bladder. The following previously reported hypotheses were evaluated: (1) aldosterone stimulates sodium transport by increasing the amount of sodium available to the sodium pump; (2) aldosterone enhances energy produc...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.64.1.330

    authors: Goodman DB,Allen JE,Rasmussen H

    更新日期:1969-09-01 00:00:00

  • neurogenin2 elicits the genesis of retinal neurons from cultures of nonneural cells.

    abstract::neurogenin2 (ngn2) encodes a basic helix-loop-helix transcription factor and plays an important role in neurogenesis from migratory neural crest cells. Its role in retinal development is poorly understood. We observed that in the developing chick retina, ngn2 was expressed in a subpopulation of proliferating progenito...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.261455698

    authors: Yan RT,Ma WX,Wang SZ

    更新日期:2001-12-18 00:00:00

  • Heparin-binding growth factor type 1 (acidic fibroblast growth factor): a potential biphasic autocrine and paracrine regulator of hepatocyte regeneration.

    abstract::Heparin-binding growth factor type 1 (HBGF-1; sometimes termed acidic fibroblast growth factor) is potentially an important factor in liver regeneration. HBGF-1 alone (half-maximal effect at 60 pM) stimulated hepatocyte DNA synthesis and bound to a high-affinity receptor (Kd = 62 pM; 5000 per cell). Epidermal growth f...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.86.19.7432

    authors: Kan M,Huang JS,Mansson PE,Yasumitsu H,Carr B,McKeehan WL

    更新日期:1989-10-01 00:00:00

  • Metabolic connectivity mapping reveals effective connectivity in the resting human brain.

    abstract::Directionality of signaling among brain regions provides essential information about human cognition and disease states. Assessing such effective connectivity (EC) across brain states using functional magnetic resonance imaging (fMRI) alone has proven difficult, however. We propose a novel measure of EC, termed metabo...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1513752113

    authors: Riedl V,Utz L,Castrillón G,Grimmer T,Rauschecker JP,Ploner M,Friston KJ,Drzezga A,Sorg C

    更新日期:2016-01-12 00:00:00

  • Gas uptake and chemical aging of semisolid organic aerosol particles.

    abstract::Organic substances can adopt an amorphous solid or semisolid state, influencing the rate of heterogeneous reactions and multiphase processes in atmospheric aerosols. Here we demonstrate how molecular diffusion in the condensed phase affects the gas uptake and chemical transformation of semisolid organic particles. Flo...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1103045108

    authors: Shiraiwa M,Ammann M,Koop T,Pöschl U

    更新日期:2011-07-05 00:00:00

  • Bystander effect produced by radiolabeled tumor cells in vivo.

    abstract::The bystander effect, originating from cells irradiated in vitro, describes the biologic response(s) of surrounding cells not directly targeted by a radiation insult. To overcome the limitations of in vitro tissue culture models and determine whether a bystander effect that is initiated by the in vivo decay of a radio...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.182209699

    authors: Xue LY,Butler NJ,Makrigiorgos GM,Adelstein SJ,Kassis AI

    更新日期:2002-10-15 00:00:00

  • Bombesin stimulates insulin secretion by a pancreatic islet cell line.

    abstract::The amphibian tetradecapeptide, bombesin (BBS) has been shown to stimulate insulin secretion both in vivo and by pancreatic islet cells in vitro. To determine whether BBS can act directly on pancreatic beta cells, we examined its effects on insulin secretion by HIT-T15 cells (HIT cells), a clonal islet cell line. Addi...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.81.6.1822

    authors: Swope SL,Schonbrunn A

    更新日期:1984-03-01 00:00:00

  • Nuclear receptor RORα regulates pathologic retinal angiogenesis by modulating SOCS3-dependent inflammation.

    abstract::Pathologic ocular angiogenesis is a leading cause of blindness, influenced by both dysregulated lipid metabolism and inflammation. Retinoic-acid-receptor-related orphan receptor alpha (RORα) is a lipid-sensing nuclear receptor with diverse biologic function including regulation of lipid metabolism and inflammation; ho...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1504387112

    authors: Sun Y,Liu CH,SanGiovanni JP,Evans LP,Tian KT,Zhang B,Stahl A,Pu WT,Kamenecka TM,Solt LA,Chen J

    更新日期:2015-08-18 00:00:00

  • Mapping of linear and circular forms of mouse mammary tumor virus DNA with restriction endonucleases: evidence for a large specific deletion occurring at high frequency during circularization.

    abstract::Rat hepatoma cells infected with mouse mammary tumor virus contain multiple forms of unintegrated viral DNA when grown in the presence of glucocorticoids. Using the DNA transfer procedure of Southern, we have prepared restriction endonuclease fragment maps of these forms of viral DNA. The maps indicate that: (i) the m...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.75.5.2112

    authors: Shank PR,Cohen JC,Varmus HE,Yamamoto KR,Ringold GM

    更新日期:1978-05-01 00:00:00

  • Designing a functional type 2 copper center that has nitrite reductase activity within α-helical coiled coils.

    abstract::One of the ultimate objectives of de novo protein design is to realize systems capable of catalyzing redox reactions on substrates. This goal is challenging as redox-active proteins require design considerations for both the reduced and oxidized states of the protein. In this paper, we describe the spectroscopic chara...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1212893110

    authors: Tegoni M,Yu F,Bersellini M,Penner-Hahn JE,Pecoraro VL

    更新日期:2012-12-26 00:00:00

  • Vitamin D-mediated modifications in protein-DNA interactions at two promoter elements of the osteocalcin gene.

    abstract::By the combined use of DNase I footprinting, electrophoretic mobility-shift assay, and methylation interference analysis, we have identified a series of sequence-specific protein-DNA interactions in the 5' flanking region of the rat osteocalcin gene. Stimulation of osteocalcin gene expression by 1,25-dihydroxyvitamin ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.87.5.1701

    authors: Markose ER,Stein JL,Stein GS,Lian JB

    更新日期:1990-03-01 00:00:00

  • The evolutionary dynamics of the Saccharomyces cerevisiae protein interaction network after duplication.

    abstract::Gene duplication is an important mechanism in the evolution of protein interaction networks. Duplications are followed by the gain and loss of interactions, rewiring the network at some unknown rate. Because rewiring is likely to change the distribution of network motifs within the duplicated interaction set, it shoul...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0707293105

    authors: Presser A,Elowitz MB,Kellis M,Kishony R

    更新日期:2008-01-22 00:00:00

  • Comparative mapping of Andropogoneae: Saccharum L. (sugarcane) and its relation to sorghum and maize.

    abstract::Comparative genetic maps of Papuan Saccharum officinarum L. (2n = 80) and S. robustum (2n = 80) were constructed by using single-dose DNA markers (SDMs). SDM-framework maps of S. officinarum and S. robustum were compared with genetic maps of sorghum and maize by way of anchor restriction fragment length polymorphism p...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.94.26.14261

    authors: Guimarães CT,Sills GR,Sobral BW

    更新日期:1997-12-23 00:00:00

  • Monoclonal antibodies against human Ia antigens stimulate monocytes to secrete interleukin 1.

    abstract::The monoclonal antibodies (mAb) DA6.147, DA6.164, and HIG.48 against human Ia antigens, but not the W6/32 mAb against human class I major histocompatibility complex antigens or the anti-monocyte OKM1 and 63D3 mAb, stimulated monocytes to secrete interleukin 1 (IL-1). IL-1 was measured by its property of promoting the ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.82.19.6652

    authors: Palacios R

    更新日期:1985-10-01 00:00:00

  • Seasonally fluctuating selection can maintain polymorphism at many loci via segregation lift.

    abstract::Most natural populations are affected by seasonal changes in temperature, rainfall, or resource availability. Seasonally fluctuating selection could potentially make a large contribution to maintaining genetic polymorphism in populations. However, previous theory suggests that the conditions for multilocus polymorphis...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1702994114

    authors: Wittmann MJ,Bergland AO,Feldman MW,Schmidt PS,Petrov DA

    更新日期:2017-11-14 00:00:00

  • Photoregulation of biological activity by photocromic reagents. II. Inhibitors of acetylcholinesterase.

    abstract::The enzymic activity of acetylcholinesterase can be photoregulated through the mediation of photochromic inhibitors of the enzyme. N-p-phenylazophenyl-N-phenylcarbamyl fluoride, an irreversible inhibitor of acetylcholinesterase, exists as two geometric isomers which are interconvertible through the action of light. Th...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.64.3.1103

    authors: Bieth J,Vratsanos SM,Wassermann N,Erlanger BF

    更新日期:1969-11-01 00:00:00