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Current Opinion in Biotechnology
Volumn 6, Issue 1, 1995, Pages 50-58
Marker proteins for gene expression
Author keywords

[No Author keywords available]
Indexed keywords

MARKER; PROTEIN;
EID: 0028936408     PISSN: 09581669     EISSN: None     Source Type: Journal    
DOI: 10.1016/0958-1669(95)80009-3     Document Type: Article
Times cited : (100)
References (50)
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    • of outstanding interest, Describes the use of the Aequorea victoria GFP as a marker. Expression of the cDNA for the GFP yields a fluorescent reporter molecule in bacteria and eurkaryotic cells that does not require exogenous substrates and co-factors. Because the fluorescent chromophore of this protein is derived through an autocatalytic mechanism, GFP expression can be used to monitor genetic activity in living organisms. This represents a new type of reporter not previously available in molecular genetic analysis.
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    • of special interest, The cardiac myosin light-chain 2v gene, MLC-2v, has served as a model system to identify the pathways that restrict the expression of cardiac muscle genes to particular chambers of the heart during cardiogenesis. In this paper, to elucidate the mechanisms of genetic regulation in the MLC-2v promoter, transgenic mice are generated that harbor mutations in five distinct cis-regulatory elements. The effects of the mutations are ascertained by expression of firefly luciferase coupled to the promoter.
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    • Tetracycline-regulated cardiac gene expression in vivo
    • of special interest, A chimeric transactivator, designated tetracycline-controlled transactivator, is used to demonstrate gene regulation in vivo in rat cardiac muscle. The firefly luciferase gene coupled to a tet operator is injected directly into the cardiac muscle, and gene regulation is monitored with oral doses of tetracycline. Luciferase activity can be controlled over two orders of magnitude.
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    • of special interest, β-galactosidase and CAT activity are used together to analyze a bidirectional promoter in Drosophila melanogaster. Mutations within the bidirectional promoter abolish expression from both genes.
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    • of special interest, Transgenic mice containing the CAT gene coupled to promoter for 3-hydroxy-3-methylglutaryl-CoA (HMG) reductase have previously been shown to ubiquitously express reporter activity. In this paper, a similar construct coupling the HMG promoter to the β-galactosidase gene fails to yield similar results. The authors suggest that the β-galactosidase gene may exert a cis effect on expression in transgenic mice.
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    • of special interest, These authors exploit β-galactosidase and firefly luciferase expression to develop a method for introducing genes into chick embryos by lipospermadine-based transfection. The cationic lipid Transfectam™ was used to transfect the reporter gene generally, or to target the gene locally through microinjection. Quantitative analysis of the method is carried out using luciferase; β-galactosidase is used for determination of spatial expression.
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    • of outstanding interest, Compartmentalization of gene expression during sporulation in Bacillus subtilis is studied using the β-galactosidase reporter gene. Expression of β-galactosidase activity is visualized by fluorescence microscopy using a fluorogenic substrate. Through the use of different promoters, gene expression can be clearly seen either throughout the cell, in the prespore region, and in the forespore.
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    • of special interest, Different stably transformed cell lines are constructed, containing the firefly luciferase gene coupled to an estrogen response element, a retinoid response element, or a 12-O-tetradecanoylphorbol-13-acetate-responsive element. Cell lines are selected by examining the induction of luminescence by the various effectors in individual cells using an intensified CCD camera to detect photon emission. After detection, the selected cells are repeatedly subcultured until a pure clonal culture is obtained.
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    • A novel circadian phenotype based on firefly luciferase expression in transgenic plants
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    • DnaK, DnaJ and GrpE form a cellular chaperone machinery capable of repairing heat-induced protein damage
    • of outstanding interest, The actions of DnaK (Hsp70), DnaJ, and GrpE in the molecular chaperone mechanism are investigated both in vivo and in vitro using firefly luciferase as a model protein. In E. coli cells inhibited for protein synthesis by antibiotics, luciferase can be thermally denatured at 42°C and then renatured at 30°C (with a 50% recovery of activity). This renaturation requires expression of DnaK, DnaJ, and GrpE from host genes; however, the presence of these genes does not protect against the thermal denaturation. Analogous results are obtained in vitro using purified components. This paper is a good example of a dynamic analysis through reporter activity both in living cells and in a reconstituted cell-free system.
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    • Inhibition of translational initiation in Saccharomyces cerevisiae by secondary structure: the roles of the stability and position of stem-loops in the mRNA leader
    • of special interest, To study the effects of stem-loop structures in mRNA translation, modular gene-expression systems are developed using the CAT or firefly luciferase reporter genes. After correction for changes in transcription, translational efficiency in yeast cells is found to be related to the predicted stability of the stem-loop structures and by their position in the 5' UTR. Results obtained using either of the reporter genes are similar.
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    • Oliveira1    Van den Heuvel2    McCarthy3
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    • Translational repression by the human iron-regulatory factor (IRF) in Saccharomyces cerevisiae
    • of special interest, The regulation of the synthesis of ferritin and erythroid 5-aminolevulinate synthase in mammalian cells is mediated by the interaction of the IRF with a specific recognition site, the IRE, in the 5' UTRs of the respective mRNAs. In this paper, this regulation mechanism is investigated both in yeast cells and in cell-free extracts using human IRF and firefly luciferase coupled to an IRE. Changes in expression of luminescence are correlated to changes in protein synthesis using antibodies to luciferase.
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    • The role of the 3'-untranslated region of non-polyadenylated plant viral mRNAs in regulating translational efficiency
    • of special interest, The genome of positive-sense RNA plant viruses functions as an mRNA but is not polyadenylated. In this paper, the role of the 3' UTRs of several viral genomes is explored using fusions to reporter genes coding for β-glucuronidase and firefly luciferase. Using electroporation of in vitro synthesized RNA constructs, differences in mRNA stability and translation efficiency are measured.
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    • The intron of Arabidopsis thaliana polyubiquitin genes is conserved in location and is a quantitative determinant of chimeric gene expression
    • of special interest, Polyubiquitin genes in Arabidopsis contain an intron in the 5' UTR immediately upstream of the initiator methionine codon. This report employs β-glucuronidase and luciferase activity to analyze the effect of these introns on expression.
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    • The sequences of and distance between two cis-acting signals determine the efficiency of ribosomal frameshifting in human immunodeficiency virus type 1 and human T-cell leukemia virus type II in vivo
    • of outstanding interest, Sequences in HTLV-2 induce translational frameshifting to overcome the termination codon at the end of the gag gene. In this paper, these sequences are inserted between the β-galactosidase and firefly luciferase genes. Without frameshifting, only β-galactosidase activity is detectable; with frameshifting, a β-galactosidase-luciferase fusion is synthesized that exhibits luminescence activity. The efficiency of frameshifting is determined as the ratio of luminescence to β-galactosidase activity. Expression of β-galactosidase and luciferase are correlated with immunoreactivity in western blots.
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    • Evidence that phosphorylation events participate in thyroid hormone action.
    • of outstanding interest, The role of phosphorylation in signal transduction by thyroid hormone receptor is investigated using luciferase coupled to thyroid hormone response elements. Results show that stimulation of luminescence by thyroid hormone T3 is enhanced by addition of a protein phosphatase inhibitor, okadaic acid. The addition of okadaic acid without T3 has no effect on gene expression. Similarly, simulation by T3 is diminished by addition of a protein kinase inhibitor, H7. Again, without T3, the inhibitor has no effect.
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    • A temperature-sensitive mutant of human p53
    • of special interest, The activity of a temperature-sensitive mutant of p53 is analyzed using firefly luciferase coupled to p53-binding elements. Mutations of p53 are commonly associated with human cancers.
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    • Zhang1    Guo2    Hu3    Liu4    Shay5    Deisseroth6
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    • The use of bacterial luciferase for monitoring the environmental regulation of expression of genes encoding virulence factors in Listeria monocytogenes
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    • Characterization of denatured protein inducers of the heat shock (stress) response in Xenopus laevis oocytes
    • of special interest, Stress response in cells is believed to be induced by intracellular accumulation of denatured proteins. This paper studies the stress response in Xenopus laevis oocytes by injection of denatured protein derivatives. Cellular stress is measured by induction of β-galactosidase activity from a heat-shock promoter. The results show that stress response is dependent on the mode of protein denaturation and the location of intracellular injection.
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    • Use of reporter genes for the isolation and characterisation of different classes of sporulation mutants in the yeast Saccharomyces cerevisiae
    • of special interest, Sporulation in yeast is not readily observable for lack of clear morphological changes, thus making mutants in the developmental pathways difficult to isolate. In this paper, the β-galactosidase reporter gene is coupled to a sporulation-specific promoter to provide a clear phenotype during the sporulation process. From this phenotype, three classes of sporulation mutations are isolated: those which overexpress the reporter gene under sporulation conditions, those which do not express the gene under any condition, and those which express the gene in vegetative cells not undergoing sporulation.
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    • Functional testing of human dopamine D1 and D5 receptors expressed in stable cAMP-responsive luciferase reporter cell lines
    • of outstanding interest, A good example showing the ability to assay receptor binding using reporter genes. The firefly luciferase gene is coupled to several CREs in a stably transformed cell line. Subsequent transfection with genes encoding human dopamine D1 and D5 receptors causes dose-dependent modulation of luminescence to dopamine agonists and antagonists. The rank of potency of the agonists and antagonists corresponds to published receptor-binding data.
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    • Functional coupling of human adenosine receptors to a ligand-dependent reporter gene system
    • of outstanding interest, The cAMP-responsive cell line described in [23] for the analysis of dopamine receptors, is used for the analysis of human adenosine receptors (A1, A2a, and A2b). The intent and results of this study are analogous to those of [23], demonstrating the ability to analyze different classes of G-coupled receptors using a common indicator cell line to reveal changes in intracellular cAMP.
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    • Castanon1    Spevak2
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    • Luminescence luteinizing hormone/choriogonadotropin (LH/CG) bioassay: measurement of serum bioactive LH/CG during early pregnancy in human and macaque
    • of special interest, A bioluminescent LH/CG bioassay is developed using the activation of firefly luciferase expression in cells expressing a human LH/CG receptor cDNA. This assay indicates hormone bioactivity, which cannot be reliably ascertained by immunoreactivity. The luciferase gene, driven from a cAMP-dependent promoter construct, yields a dose-dependent response to human LH or CG. Treatment with follicle-stimulating hormone, thyroid-stimulating hormone, prolactin, growth hormone, adrenocorticotropin, insulin, prostaglandins, and several neurotransmitters has no effect. Stimulation of luminescence is observed, however, in the presence of basic fibroblast growth factor.
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    • Jia1    Perlas2    Su3    Moran4    Lasley5    Ny6    Hsueh7
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    • An assay for transforming growth factor-β using cells transfected with a plasminogen activator inhibitor-1 promoter-luciferase construct
    • of special interest, TGF-β is a potent regulator of cellular differentiation, proliferation, migration, and protein expression. In this paper, a bioassay for TGF-β is developed using stably transformed cells containing the firefly luciferase gene coupled to a plasminogen activator inhibitor-1 promoter. The cell line yields dose-dependent luminescence to TGF-β in the range of 0.2 mM to 〉 30 mM, providing greater sensitivity and specificity than widely used alternative bioassays.
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    • Abe1    Harpel2    Metz3    Nunes4    Loskutoff5    Rifkin6
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    • Development of specific bioluminescent in vitro assays for selecting potential antimineralocorticoids
    • of special interest, A ‘minimal’ chimeric receptor is constructed to analyze the biological activity of various antimineralocorticoids. Transient and stable cell lines containing the firefly luciferase gene are used to indicate steroid binding to the receptor.
    • (1994) J Steroid Biochem Mol Biol , vol.49 , pp. 31-38
    • Jausons-Loffreda1    Balaguer2    Auzou3    Pons4
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    • The adenovirus-mediated delivery of a reporter gene permits the assessment of androgen receptor function in genital skin fibroblast cultures
    • of outstanding interest, This is an interesting example of the use of a reporter gene to diagnose a clinical condition. Defects in the androgen receptor cause a spectrum of abnormalities in male carriers, ranging from feminized phenotype to minor defects in fertility. To evaluate receptor functionality, fibroblasts cultured from genital skin are infected with recombinant adenovirus to deliver an androgen-inducible firefly luciferase gene. In fibroblasts from normal individuals, androgen causes an 11- to 200-fold increase in luminescence, corresponding to the level of androgen receptor detected in ligand-binding assays. In contrast, only a negligible increase (about 1.2-fold) is evident in fibroblasts from men with testicular feminization.
    • (1993) J Biol Chem , vol.268 , pp. 26063-26066
    • McPhaul1    Deslypere2    Allman3    Gerard4
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    • Optical biosensor for environmental on-line monitoring of naphthalene and salicylate bioavailability with an immobilized bioluminescent catabolic reporter bacterium
    • of special interest, Sensors for continuous on-line monitoring of naphthalene and salicylate bioavailability are developed from Pseudomonas fluorescens containing a bacterial luciferase operon coupled to the nahG promoter. The engineered cells are immobilized onto an optical light guide, and the probe is then inserted into a measurement cell, which receives a waist stream mixed with maintenance medium. Reproducible luminescent signals are achieved with repeated exposures to naphthalene or salicylate. The sensor is also tested with jet fuel and leachate from contaminated soil, both of which contain naphthalene.
    • (1994) Appl Environ Microbiol , vol.60 , pp. 1487-1494
    • Heitzer1    Malachowsky2    Thonnard3    Bienkowski4    White5    Sayler6
  • 30
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    • Rapid and sensitive pollutant detection by induction of heat shock gene-bioluminescence gene fusions
    • of special interest, An example of the use of engineered bacteria as environmental sensors. An E. coli strain is employed that contains a bacterial luciferase operon from Vibrio fischeri. The luciferase operon is coupled to two heat-shock promoters, dnaK and grpE, to indicate the presence of dissolved metals, solvents, crop-protection chemicals, and other organic compounds. Photon production is non-invasive because the entire luminescence operon is used.
    • (1994) Appl Environ Microbiol , vol.60 , pp. 414-1420
    • Van Dyk1    Majarian2    Konstantinov3    Young4    Dhurjati5    LaRosa6
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    • Detection of herpes simplex virus by measurement of luciferase activity in an infected-cell lysate
    • of outstanding interest, A stably transformed cell line is developed that expresses high levels of luciferase activity following infection with herpes simplex virus. The cell line contains a herpes simplex virus type 1 promoter-luciferase chimeric gene, which yields a greater than 10 000-fold increase in luminescence upon virus infection. This paper shows the utility of firefly luciferase as an indicator of specific viral activity and also the wide assay range required in some reporter applications.
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    • Olivo1
  • 32
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    • A cell line that expresses a reporter gene in response to infection by Sindbis virus: a prototype for detection of positive strand RNA viruses
    • of special interest, Describes the development of a stably transformed cell line that contains a defective Sindbis virus genome under control of a Rous sarcoma virus promoter and the luciferase gene downstream of the viral subgenomic RNA promoter. The cell line expresses high levels of luciferase activity following infection with Sindbis virus and related variant viruses.
    • (1994) Virology , vol.198 , pp. 381-384
    • Olivo1    Frolov2    Schlesinger3
  • 33
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    • Tau-β-galactosidase, an axon-targeted fusion protein
    • of special interest, As a genetic marker of neuronal cells, β-galactosidase is limited by its inability to readily diffuse into axons. In this paper, a modified form of the enzyme is constructed by fusing the cDNA encoding bovine microtubule-binding protein, tau, onto the β-galactosidase gene. Using an enhancer-trap transposon in Drosophila, this modified reporter is used to mark various neuronal cell types, as well as muscle fibers and glial cells.
    • (1994) Proc Natl Acad Sci USA , vol.91 , pp. 5972-5976
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    • Distinct modes of human immunodeficiency virus type 1 proviral latency revealed by superinfection of nonproductively infected cell lines with recombinant luciferase-encoding viruses
    • of special interest, To study mechanisms of cellular latency in HIV infection, a recombinant HIV is constructed with firefly luciferase replacing the nef gene. This recombinant virus is used to rapidly measure active viral growth on different latent cell lines. The study reveals both cis and trans mechanisms for latency.
    • (1994) J Virol , vol.68 , pp. 654-660
    • Chen1    Saksela2    Andino3    Baltimore4
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    • Molecular marker systems for detection of genetically engineered micro-organisms in the environment
    • of outstanding interest, This review covers the use of reporter genes as markers for the detection of genetically engineered cells in the environment. Coverage is given to antibiotic resistance genes, β-galactosidase, catechol 2,3-dioxygenase (encoded by xylE), 2,4-dichlorophenoxyacetate monooxygenases, and bacterial luciferases. The use of bacterial luciferases constitutes the greatest part of the review. Detection methods for luminescence are also discussed.
    • (1994) Microbiology , vol.140 , pp. 5-17
    • Prosser1
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    • Gene transfer into mammalian cells by particle bombardment
    • of special interest, Examines the factors affecting the efficient transfer of genes into mammalian cells by particle bombardment. Firefly luciferase is used for quantitative analysis of gene introduction, and β-galactosidase is used to visualize the spatial pattern of particles in cell-culture dishes. The particle bombardment method is compared with transformation by electroporation, lipofection, and diethylaminoethyl dextran.
    • (1994) Anal Biochem , vol.217 , pp. 185-196
    • Heiser1
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    • Direct myocardial transfection in two animal modes
    • of special interest, The effectiveness of gene therapy through direct injection of DNA into the myocardium is investigated using the firefly luciferase gene. The effect of the amount of DNA carried by the delivery vehicle and its volume were quantitatively analyzed. Also examined are the persistence of foreign gene expression and the feasibility of a percutaneous injection method.
    • (1993) Lab Invest , vol.68 , pp. 18-25
    • Gal1    Weir2    Leclerc3    Pickering4    Hogan5    Isner6
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    • Coronary restenosis and gene therapy
    • of special interest, The long-term effectiveness of coronary angioplasty is limited by the proliferative response of vascular smooth muscle cells to the site of vascular injury imposed by the technique. Currently, non-permanent gene therapy at the site of vascular injury is being investigated as a means of locally inhibiting the proliferative response. This paper evaluates the utility of an adenovirus vector for introducing DNA into vascular tissue, both in vivo and in vitro, using the firefly luciferase and β-galactosidase reporter genes.
    • (1994) Texas Heart Inst J , vol.21 , pp. 104-111
    • Mazur1    Ali2    Raizner3    French4
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    • 3H]acetylCoA concentration
    • 3H]acetyl-CoA from the reaction. The new method yields several fold greater sensitivity than the conventional method, but it is still estimated to be 14-fold less sensitive than the assay for firefly luciferase.
    • (1994) Anal Biochem , vol.219 , pp. 147-153
    • Chireux1    Raynal2    Weber3
  • 41
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    • Gene expression of a thermostable β-galactosidase in mammalian cells and its application in assays of eukaryotic promoter activity
    • of special interest, Initial experiments in the use of thermostable β-galactosidase as a reporter gene are described. The thermostable reporter is cloned from the thermoacidophilic archaebacterium Sulfolobus solfataricus. The reporter exhibits very little activity at 37°C; enzyme activity is assayed by incubation at 75°C. A preliminary comparison with CAT is also given.
    • (1994) Biotechnol Appl Biochem , vol.19 , pp. 233-244
    • Cannio1    dePascale2    Rossi3    Bartolucci4
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    • Selective inactivation of eukaryotic β-galactosidase in assays for inhibitors of HIV-1 TAT using bacterial β-galactosidase as a reporter enzyme
    • of special interest, Describes a method for increasing the sensitivity of the β-galactosidase assay by reducing interference from endogenous enzymatic activity. Heat treatment at 50°C for 1 h inactivates the β-galactosidase activity endogenous to several eukaryotic cell lines by as much as 40-fold without adversely affecting the activity of bacterial β-galactosidase.
    • (1993) Anal Biochem , vol.215 , pp. 24-30
    • Young1    Kingsley2    Ryan3    Dutko4
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    • Chemiluminescent reporter gene assays: sensitive detection of the GUS and SEAP gene products
    • of special interest, Describes chemiluminescent reporter gene assays for human placental SEAP and β-glucuronidase using adamantyl dioxetane derivatives.
    • (1994) Biotechniques , vol.17 , pp. 172-177
    • Bronstein1    Fortin2    Voyta3    Juo4    Edwards5    Olesen6    Lijam7    Kricka8
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    • Chemiluminescent and bioluminescent reporter gene assays
    • of outstanding interest, This review describes the available reporter genes assayed by bioluminescence and chemiluminescence. It provides coverage of several luciferases and photoproteins; cheluminescent methods are limited to adamantyl dioxetane chemistries. Some comparative information is given, together with a brief discussion of various detection methods.
    • (1994) Anal Biochem , vol.219 , pp. 169-181
    • Bronstein1    Fortin2    Stanley3    Stewart4    Kricka5
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    • Modulation of firefly luciferase stability and impact on studies of gene regulation
    • (1991) Gene , vol.103 , pp. 171-177
    • Thompson1    Hayes2    Lloyd3
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    • Promoter independent down-regulation of the firefly luciferase gene by T3 and T3 receptor in CV1 cells
    • of outstanding interest, Down-regulation of luciferase expression is demonstrated in CV-1 cells expressing T3 receptor in the presence of T3. Down-regulation is shown to be caused by the gene itself and not by other vector sequences. The mechanism underlying this effect is not identified and it may not be general to other cell types. Even so, the results show the general importance of confirming that reporters behave as expected in specific host cells under specific experimental conditions. A ‘baseline’ genetic construction should, thererfore, be included in most quantitative analyses using genetic reports.
    • (1993) Mol Cell Endocrinol , vol.95 , pp. 101-109
    • Tillman1    Crone2    Kim3    Sprung4    Spindler5
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    • Intracellular generation of superoxide as a by-product of Vibrio harveyi luciferase expressed in Escherichia coli
    • of outstanding interest, In a study of the E. coli soxRS regulon, which regulates various antioxidant defense enzymes, bacterial luciferase is found to cause gene induction in the absence of superoxide-generating agents. It is postulated that without aldehyde substrate the bacterial luciferase could generate superoxide through autoxidation of the flavin. This is supported by biochemical and genetic data. This observation may be general for all bacterial luciferases, but does not apply to other luciferases.
    • (1994) J Bacteriol , vol.176 , pp. 2293-2299
    • Gonzalez-Flecha1    Demple2
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    • Use of transcriptional fusions to monitor gene expression: a cautionary tale
    • of outstanding interest, Bacterial luciferase yields anomalous results in E. coli and Salmonella typhimurium when coupled to the leu-500 promoter. In contrast, coupling to CAT yields results consistent with those previously reported using galactokinase. Promoter activities are assayed from single-copy insertions into the host (S. typhimurium) chromosome and from low-copy and multiple-copy plasmids (in E. coli and S. typhimurium). Anomalous results are also achieved when bacterial luciferase is coupled to the proU promoter in E. coli, whereas β-galactosidase yields expected results. Coupling of bacterial luciferase to the lac promoter or the gyrB promoter does, however, report expected gene expression patterns.
    • (1994) J Bacteriol , vol.176 , pp. 2132-2182
    • Forsberg1    Pravitt2    Higgins3
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    • Firefly luciferase engineered for improved genetic reporting
    • of outstanding interest, This brief communication describes a new form of the firefly luciferase gene engineered for more reliable use as a genetic reporter. The primary modification yields a cytoplasmic form of the luciferase, which may be important to avoid the disruption of normal peroxisomal function in exogenous hosts. Other modifications make the gene more convenient to use and minimize potential interference in diverse biological systems.
    • (1995) Promega Notes , vol.49 , pp. 14-21
    • Sherf1    Wood2

* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.