real time reverse transcription polymerase chain reaction array
Role of promyelocytic leukemia (PML) and small ubiquitin like modifier (SUMO) proteins in alternative lengthening of telomeres
... 3.11 Components of PCR reaction for addition of FLAG tag to PML C/C50 Table 3.12 PCR reaction conditions for addition of FLAG tag to PML C/C- 51 Table 3.13 Restriction digest reaction mix 51 Table ... DNA sequencing reaction mix 54 Table 3.15 PCR reaction conditions for DNA sequencing 54 Table 3.16 Solutions for Western blot 60 Table 3.17 Crystal violet solution 63 Table 3.18 Reaction mix of ... cases, SUMOylation of transcription factors represses their transcriptional activity Thus when the SUMOylation sites are mutated, there is generally hyperactivation of transcription There are...
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Báo cáo y học: "Cell Cycle Arrest by a Natural Product via G2/M Checkpoint"
... activate the transcription of a variety of apoptosis-associated genes, and to program cell death in response to genotoxic stresses [16, 17] 14-3-3σ (stratifin), p21, and Gadd45 are three major transcriptional ... CKBM on cell cycle was evaluated using flow cytometric analysis We observed a dose-dependent but timeindependent effect of CKBM on the cell cycle After 72 hr of % CKBM treatment, cells in the G2/M...
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Báo cáo y học: "Comment on and reply to "Analysis of variation of amplitudes in cell cycle gene expression" by Liu, Gaido and Wolfinger: On the analysis of gene expression during the normal, eukaryotic, cell cycle" docx
... period of time covering a significant fraction of the interdivision time 2) The rise in cell numbers during division should occur over a relatively small fraction of the total interdivision time It ... (Minimum Information About a Microarray Experiment) or MAGE (microarray gene expression object model) standards, so that the public data can be analyzed 18) Microarray results should be compared ... the second cycle 5) Successive generations (i.e the time between rises in cell number) should be of equal length and equal to the doubling time of cells in exponential growth 6) Data points should...
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Báo cáo khoa học: Bridging the gap between in silico and cell-based analysis of the nuclear factor-jB signaling pathway by in vitro studies of IKK2 ppt
... control rhIKK2 in vitro kinase assay with no GST-IjBa substrate The time (in minutes) on the abscissa indicates the time the reactions were stopped with trichloroacetic acid and the plot shows ... NF-jB in the cell 1686 Recombinant human IKK2 (rhIKK2) time- course reaction was carried out for 113 in 50 mm Tris HCl, pH 7.5, and 10 mm MgCl2 Reactions were performed in a nal volume of 45 lL (15 ... assay and the negative control was stopped at time zero Reaction sample (40 lLặwell)1) was precipitated with 40 lL of 12% w v trichloroacetic acid Reactions were performed using the same protocol...
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Báo cáo khoa học: Optimization of an Escherichia coli system for cell-free synthesis of selectively 15N-labelled proteins for rapid analysis by NMR spectroscopy pdf
... the cross-peaks from the side -chain amide groups (backbone and side -chain NH groups were labelled in the amino acid used in the reaction) The cross-peak from the side -chain NH of W121 is labelled ... following issues were addressed: (a) how yields compare, when transcription is performed by T7 RNA polymerase from a T7 promoter or by E coli RNA polymerase from tandem phage k pR and pL promoters; (b) ... RNA polymerase performs as well as the E coli holoenzyme in the in vitro coupled transcription/ translation system As many modern plasmid vectors used for protein overproduction are based on transcription...
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Báo cáo khoa học: Analysis of the contribution of changes in mRNA stability to the changes in steady-state levels of cyclin mRNA in the mammalian cell cycle doc
... membranes were stripped twice for 15 in 250 mL of boiling 0.1 · NaCl ⁄ Cit, 0.5% SDS Reverse transcription and real- time PCR To avoid amplification of residual genomic DNA, firstly this was removed ... set for real- time PCR we always verified that there was no significant amplification product in the absence of reverse transcriptase First-strand cDNA was synthesized using lg RNA in a 20-lL reverse ... different elutriated cell fractions Transcription was inhibited with 5,6-dichloro-1-b-d-ribofuranosylbenzimidazole (DRB) and mRNA measured at 0, 30, 60, 120 and 180 by real- time PCR (Fig 3) Half-life...
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báo cáo hóa học:" ApoG2 induces cell cycle arrest of nasopharyngeal carcinoma cells by suppressing the c-Myc signaling pathway" pot
... CNE-2 cells with 5, 10 and 20 μM ApoG2 for 24, 48 and 72 h This treatment resulted in dose- and timedependent inhibition of cell proliferation (Fig 1B) At 10 and 20 μM, ApoG2 inhibited about 60% ... induced cell cycle arrest at the DNA synthesis (S) phase in a large percentage of NPC cells at this time More than 60% of C666-1, CNE-1 and CNE-2 cells were arrested at S phase at 48 h after exposure ... 24 h and gradually returned to low level at 72 h since most of the CNE-2 cells were dead at this time (Fig 3B); unlike p21, expression of both cyclin D1 and cyclin E were downregulated following...
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Báo cáo hóa học: " Research Article A Robust Structural PGN Model for Control of Cell-Cycle Progression Stabilized by Negative Feedbacks" pdf
... the last instant of time Let X, with realization x, represent the state before a transition, and let Y , with realization y be the first state after that transition A Markov chain is characterized ... where N where y is a realization of a random vector with distribution p•|x An m order Markov chain which depends on the m previous instants of time is equivalent to a Markov chain whose states ... value of xi at time t is not affected for its previous value at time t − k The constant parameter pi , for the state variable xi , represents the number of previous instants of time at which the...
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Báo cáo y học: "Genome-wide analysis of the diatom cell cycle unveils a novel type of cyclins involved in environmental signaling" pptx
... intervals after illumination, samples were taken for expression analysis by real- time quantitative polymerase chain reaction (qPCR) CDKs and CDK interactors CDKs CDKs are serine/threonine kinases ... mentioned above Samples were taken for real- time qPCR after hours of incubation in the dark Real- time qPCR For RNA extraction, × 107 cells were collected at each time point, fast frozen in liquid ... normalization gene evaluation (a) Real- time qPCR cycle threshold (Ct) values of candidate housekeeping genes during a 12-h (sampling every hour) synchronization time series (b) Variation of Ct...
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báo cáo khoa học: "Regulation of cell cycle transition and induction of apoptosis in HL-60 leukemia cells by lipoic acid: role in cancer prevention and therapy" ppt
... is both time and dose dependent Initially, we investigated the effect of LA on cell growth inhibition Exponentially growing HL-60 cells were treated with increasing doses and exposure times of ... was added to the culture media to the final concentration specified in the text At the specified times, control and treated cells were harvested Cell count was performed using a hemocytometer and ... apoptosis and cell cycle arrest and the underlying mechanisms of action Our study shows for the first time that LA is capable to http://www.jhoonline.org/content/1/1/4 block multiple cell cycle checkpoints...
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Báo cáo y học: "Cell cycle G2/M arrest through an S phase-dependent mechanism by HIV-1 viral protein R." pps
... control or Adv-Vpr (MOI 1.0) and released from the block at time The cell cycle profiles measured by DNA content (a) were detected from time to 11 hours after the DT release The Cdk1-Tyr345 or Chk1Ser345 ... Synchronized G1/S HeLa cells by DT were treated with HU, UV or transduced with Adv-Vpr at time 0, collected at the indicated time, and then subjected to Western blot analysis (A) using anti-Chk1-Ser345 and ... in small but perhaps insignificant decrease of Cdc25C over time; in contrast, Vpr induced a rather strong reduction of Cdc25C over time (Figure 5A-a, second row, lanes 8-10) To ascertain Vprmediated...
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Báo cáo y học: "Analysis of variation of amplitudes in cell cycle gene expression" potx
... modeling the time- course of expression, yg(t), of a cell cycle related gene g over time t: amplitude, which we denote as Kg; duration of cycle (period), T; and phase angle, φg, which is the time in ... can be used to gauge the variation of the phase/peak expression time distribution involved in stochastic transcriptional and post-transcriptional processes for the gene in a cell population Further ... expression times for a gene within a cell population Therefore, we can use the ratio of estimated amplitudes from RPM (1) to examine the relative variability in phase/peak expression time for gene...
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Báo cáo y học: "Impairment of organ-specific T cell negative selection by diabetes susceptibility genes: genomic analysis by mRNA profiling" pot
... oligonucleotide array data via quantitative real- time PCR BMC Bioinformatics 2006, 7:23 Gene expression omnibus [http://www.ncbi.nlm.nih.gov/geo/] Quackenbush J: Computational analysis of microarray data ... to Affymetrix GeneChip 430A arrays (Santa Clara, CA, USA) Microarray data processing The Affymetrix 430A chips were scanned using standard Affymetrix protocols All arrays passed routine quality ... strains and microarray gene expression analyses to study a complex trait: the NOD model of type diabetes Genome Res 2002, 12:232-243 DeRyckere D, Mann DL, DeGregori J: Characterization of transcriptional...
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cell cycle regulation - Ebook USA ( biotechnology)
... inhibiting members of the E2F transcription factor family (Bartek et al 1996) E2Fs associate with Dp1 or Dp2 to form an active transcription factor complex upregulating a wide array of gene products ... during transcription The positive correlation between origin firing and transcriptional activity in higher eukaryotes suggests that the more open chromatin structure not only allows efficient transcription, ... clinical trials without knowing the potential outcome This is a time- consuming and risky procedure, which has been going on for too long a time The future of cell cycle research is bright and the results...
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Báo cáo y học: "BRCA1 May Modulate Neuronal Cell Cycle Re-Entry in Alzheimer Disease"
... Evidence for a transcriptional activation function of BRCA1 C-terminal region Proc Natl Acad Sci U S A 1996; 93: 13595-9 Gowen LC, Avrutskaya AV, Latour AM, et al BRCA1 required for transcription- coupled ... when problems are suspected Since the prevalence of BRCA1 increases as the disease progresses, transcription of BRCA1 may be activated early in the progression of AD This is consistent with the ... very early in the progression of the disease, long before the presence of other pathology Over time, DNA and cell cycle changes may compound, and BRCA1 and other protein expression increases,...
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Cell Cycle Regulation in Mammalian Germ Cells
... 284–286, 345, 356 tetratricopeptide repeat, 152 transcription, 166, 355, 356 transcription factors, 183–185, 188, 190, 191, 208–210, 237, 332, 333, 337 transcriptional repressors, 333 transformation, ... mitotic progression Lack of gene leads of spermatocytes arrested at metaphase-I Transcription factor important in RNA polymerase II machinery Impaired folliculogenesis is seen in knockouts leading ... architecture for cell cycle progression Transcriptional and Translational Factors Regulation of meiosis and gametogenesis also takes place at the level of transcription, which affects proliferation,...
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Modeling Cell Cycle Control and Cancer with pRB Tumor Suppressor
... carcinomas increase transcription of CCND1 via increases in β-catenin/TCF signaling resulting from loss of the APC tumor suppressor (Tetsu and McCormick 1999) Neuroblastomas increase transcription ... inhibits PU.1, a transcription factor that stimulates the macrophage lineage, upon which developing erythroblasts are dependent for enucleation (Iavarone et al 2004) The E2F/DP Transcription Factor ... members, and are not competent for activating transcription 4.1 E2F Target Genes and Repression Interaction of E2F1–3/DP complexes with pRB converts these transcriptional activators to repressor complexes...
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MouseModels of Cell Cycle Regulators - New Paradigms
... of the Rb family (Rb, p130, p107) This in turn leads to the release of E2F transcription factors and activation of transcription of E2F-responsive genes (Sherr and Roberts 1999; 2004) The D-type ... addition, Cdk4–/–Cdk6–/– MEFs proliferate well and have a similar doubling time to wild type MEFs at P2 but a significantly longer time at P4 (72 versus 52 hours) This is reflected in a drop in the percentage ... new paradigms emerging from mouse models reflect the complexity of higher mammals but at the same time prove that the molecular machinery operating the cell cycle is highly conserved and in higher...
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Senescence and Cell Cycle Control
... family transcription factors (Ohtani et al 2001) Ets-1 can directly transactivate p16INK4a and induce senescence, and this regulation is abrogated by Id-1 as a specific inhibitor Bmi-1, a transcriptional ... fate determination between senescence and immortalization These proteins bind to the E2F family transcription factors and maintain the repressor function of E2F (Hatakeyama and Weinberg 1995; ... senescenceassociated heterochromatic foci (SAHF) (Narita et al 2003), which may play a critical role in long-term transcriptional repression specific in senescent cells Interestingly, similar cell cycle reentry...
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The Retinoblastoma Gene Family in Cell Cycle Regulation and Suppression of Tumorigenesis
... Barbacid 2001; see Fig 1) Regulation of E2F Responsive Genes by pRb E2F transcription factors, named for their activity to mediate transcriptional activation of the adenovirus E2 promoter, recognize ... transition of cell cycle Whether an E2F target gene is transcriptionally activated or repressed depends on binding of pRb to E2F pRb inhibits the transcriptional activity of E2F by binding to its ... growth-inhibitory signals Finally, it was shown in a reconstitution transcription assay that chromatin is an essential component for pRb to actively repress transcription, although HDACs did not seem to play...
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