design reverse transcription and pcr for confirming gene expression patterns

... differential gene expression patterns from wild-type and ras knockout fibroblasts after serum stimulation for hour or hours Figure Overlapping of differential gene expression patterns from wild-type and ... fibroblastsreverse/N-rasexpression gene arrays.TableS6:presenceTableuseddifferentialin serum-starved ,for hours.TabledataG0-arrestedgenefibroblastsincubationfibroblasts afterin starved,-/-in forgenewithfor8forininTable ... collection and manuscript editing AN performed tasks related to maintenance and use of the animal knockout colony JdlR carried out the bioinformatics analyses of the transcriptional data ES designed and...

Ngày tải lên: 09/08/2014, 20:20

... contributions MH designed experiments, performed RNA and DNA extractions and blots, amplified and sequenced BURP gene from CS and CG genotypes, analyzed DGE data for functional categories, and drafted ... differences of Glyma04 BURP gene in Clark standard and Clark glabrous To validate the transcriptome data for the BURP gene, we performed RNA blot analysis for the Glyma04g35130 BURP gene Total RNA was ... National Center for Biotechnology Information [55] and trEMBL and Swiss prot of the European Bioinformatics Institute [56] on a Time Logic CodeQuest DeCypher Engine BURP Gene Cloning and Sequence...

Ngày tải lên: 11/08/2014, 11:21

... )1.3 A, ˚ and )10.7°, respectively, for PDwt, and 3.3 A ˚ )4.0 A and )10.6°, respectively, for PDmut Average ˚ widths of minor and major grooves are around 9.5 A ˚ in both PDwt and PDmut and 25 ... bond distances (C and E) and angles (D and F) over ns dynamics Note the strong preference for N1(A24) .N4(C7) hydrogen bond beyond 220 ps (C and D) Conformation of the PNA strand in PNAÆDNA duplexes ... cis) (A and B) and (a,e) (gauche+, trans) (C and D) conformations O6¢(G) .N2(G) hydrogen bond (C and D) is shown in dotted line (see also Fig 10 and text) Hydrogens at C2¢, C3¢, C5¢ and C8¢...

Ngày tải lên: 07/03/2014, 21:20

... expression of the gene A likely explanation for this is that the transposon-tagged psaG gene in the knock-out line used for the transformation experiments still produces a psaG transcript, and together ... wild-type and His-tagged PSI-G by high-resolution SDS ⁄ PAGE (C) Immunodetection of His and Strep tags in PSI-G constructs and transformed plants were treated separately with thermolysin and trypsin ... cloned into binary vectors under the control of the 35S promoter and terminator, and into vectors for in vitro transcription and translation An Arabidopsis line (psag-1.4 [5], termed DG in this...

Ngày tải lên: 07/03/2014, 21:20

... incubation and assayed for GO activity Protein concentration was measured using the Bradford protein assay Cloning of B subtilis GO cDNA and DNA manipulation The cloning and transformation techniques ... and BamHI/ScaI and by a PCR reaction, using the XbaI [6] and YJBr-down oligonucleotides as a primer The pT7-HisGO expression plasmid was transferred to the host BL21(DE3)pLysS E coli strain for ... cultures transformed with the pT7-GO expression vector and grown on Luria–Bertani medium, as indicated by native PAGE and GO activity staining The highest level of GO expression and GO specific...

Ngày tải lên: 08/03/2014, 16:20

... constituents and products may still be produced by such an L-form mutant, and investigated whether hitherto unknown bypass mechanisms exist This should prove useful for understanding such morphogenetic ... resulted in 75 modes in total There are 12 modes for producing lipid A and 14 for the cold-adapted form We found that six of the former 12 modes and six of the latter 14 modes produced L1-P-EtAmine ... [14–16,57] Isolation and analysis of genes of the L-form mutant strain E coli LWF1655F+ Amplification of the genes of interest (Table S6), mutation detection and analysis were essentially performed as previously...

Ngày tải lên: 22/03/2014, 21:20

... Centre for Virus Diseases of Animals, Island Riems), BT cells (ATCC CRL-1390) and MDBK cells (ATCC CCL-22) Each cell line was infected at a multiplicity of infection (m.o.i.) of and cultured for ... was performed using a FACScalibur (Becton Dickinson) Dead cells and debris were excluded by appropriate gating and 30000 events were collected Analysis was performed using CellQuestPro and Flowjo ... analyses JLD, AA and JG performed electron microscopy studies GF contributed to PBMC infection studies GF and SB coordinated the research BP and SB wrote the manuscript All authors read and approved...

Ngày tải lên: 18/06/2014, 18:20

... Centre for Virus Diseases of Animals, Island Riems), BT cells (ATCC CRL-1390) and MDBK cells (ATCC CCL-22) Each cell line was infected at a multiplicity of infection (m.o.i.) of and cultured for ... was performed using a FACScalibur (Becton Dickinson) Dead cells and debris were excluded by appropriate gating and 30000 events were collected Analysis was performed using CellQuestPro and Flowjo ... analyses JLD, AA and JG performed electron microscopy studies GF contributed to PBMC infection studies GF and SB coordinated the research BP and SB wrote the manuscript All authors read and approved...

Ngày tải lên: 20/06/2014, 01:20

... 95°C for 15 s and 60°C for Melting curve analysis was performed to identify nonspecific PCR products and primer dimers Primers were designed using Primer Express® Software for Real-Time PCR 3.0 ... was calculated for each gene The detailed qRT -PCR results for eight genes are shown in Additional file 2: Figure S2 Among 20 genes, 16 showed good correlation between qRT -PCR and microarray data ... 23 distinct regulons and their relationship to metabolism, chromosomal gene distribution and co-ordination of nuclear and plastid gene expression Gene 2005, 344:33-41 85 Khandelwal A, Elvitigala...

Ngày tải lên: 11/08/2014, 11:22

... (M) and (N), respectively Seeds in the first (most mature) pod and close to pea stock are designated as 1.1 The youngest pod and close to the pea stock is designated as 8.1 for JI 116, 8.1 for ... agriculture and horticulture When fruits and seeds undergo abscission, they provide an efficient and effective means of dispersal and propagation so that plants are maintained from generation to generation ... yield The identification and manipulation of traits and processes that influence fruit and seed dispersal are therefore of great interest in the development of strategies for crop improvement through...

Ngày tải lên: 12/08/2014, 03:20

... interpretation and in the writing HOK and HSK performed the analysis of the data and drafted the manuscript JCY and ECS participated in the design of the study and performed the statistical analysis and ... (Tables and and Figure 1) TGF-a binds to the EGF receptor with a high affinity [21] and is indispensable for the proper development of many tissues and organs, wound healing, bone resorption, and ... with log transformed G-CSF (R = -0.232, P = 0.016) and log transformed GM-CSF (R = -0.249, P = 0.009) a pathophysiological role in vascular remodeling and atherogenesis Monocytes and neutrophils,...

Ngày tải lên: 14/02/2014, 06:20

... trimer and the pentamer compared to the dimer were 34% and 30% for the wildtype, but 66% and 47% for the D392N mutant (Fig 5B) A Oligosaccharide synthesis by chitinase A Protein expression and hydrolytic ... The maximum yields of (GlcNAc)5 and (GlcNAc)6, synthesized relatively to the hydrolytic product, (GlcNAc)2, were 3% and 9% for the wild-type enzyme and 11% and 12% for the D392N mutant The synthesis ... Similar patterns were also seen with other substrates For instance, the tetramers, pentamers, and hexamers were formed during (GlcNAc)3 hydrolysis, while hexamers, heptamers, and octamers were formed...

Ngày tải lên: 20/02/2014, 01:20

... each with 2048 data points and an appropriate number of scans Zero filling and Fourier transformation led to spectra with ranges of  5400 Hz and  30 000 Hz for proton and carbon, respectively ... analysis The Kd values were 95 ± lm and 100 ± 10 lm for complexes with Glc1P and GL, respectively They were decreased seven-fold and three-fold in comparison to Kd values for corresponding complexes ... absence of ligand (d), in the presence of 20 mM arsenate (s), and in the presence of 20 mM arsenate and mM GL (.); 31P shift for the H334G mutant (,), recorded in the absence of arsenate and at only...

Ngày tải lên: 23/03/2014, 07:20

... 40, 50, 60, 70, 80, 90, and 100 mM; and CTPS concentrations were 20 lgặmL)1 for wild-type and 2024 lgặmL)1 for L109A For assays of Gln- or Gln analogue-dependent CTP formation, concentrations ... 5.0, and 10.0 mM), Gln-OH (1.0, 3.0, 5.0, 7.0, and 10 mM), or GlnNH2 (3.0, 7.0, 10.0, 15.0, and 20.0 mM) CTPS concentrations ranged between and 56 lgặmL)1 for wild-type and 554 lgặmL)1 for L109A ... NH2OH and NH2NH2 in place of NH3 [4244] Both NH2NH2 and NH2OH (and its derivatives NH2OCH3 and CH3NHOH) Exogenous NH3 and its analogues Exogenous NH3, NH2OH, and NH2NH2 all served as substrates for...

Ngày tải lên: 23/03/2014, 13:20

... performed at a low salt concentration (a requirement for CD spectroscopy) in 10 mM potassium phosphate buffer at pH 5.8 and 8.0 Materials and methods Results and discussion Protein production and ... wild-type Cel7A and pH mutant E223S/A224H/L225V/T226A/D262G at pH 5.8 and pH 8.0 and 25 °C Spectra of the wild-type Cel7A at pH 5.8 (d) and pH 8.0 (s), and the pH mutant at pH 5.8 (j) and pH 8.0 (h) ... Liljankoski, M Siika¨ aho and R Suihkonen for construction of the Cel7A mutant, and for the purification of the wild-type and the mutant protein K Rautajoki is thanked for the help with the fluorescence...

Ngày tải lên: 31/03/2014, 07:20

... interpretation and in the writing HOK and HSK performed the analysis of the data and drafted the manuscript JCY and ECS participated in the design of the study and performed the statistical analysis and ... (Tables and and Figure 1) TGF-a binds to the EGF receptor with a high affinity [21] and is indispensable for the proper development of many tissues and organs, wound healing, bone resorption, and ... with log transformed G-CSF (R = -0.232, P = 0.016) and log transformed GM-CSF (R = -0.249, P = 0.009) a pathophysiological role in vascular remodeling and atherogenesis Monocytes and neutrophils,...

Ngày tải lên: 18/06/2014, 22:20

... interpretation and in the writing HOK and HSK performed the analysis of the data and drafted the manuscript JCY and ECS participated in the design of the study and performed the statistical analysis and ... (Tables and and Figure 1) TGF-a binds to the EGF receptor with a high affinity [21] and is indispensable for the proper development of many tissues and organs, wound healing, bone resorption, and ... with log transformed G-CSF (R = -0.232, P = 0.016) and log transformed GM-CSF (R = -0.249, P = 0.009) a pathophysiological role in vascular remodeling and atherogenesis Monocytes and neutrophils,...

Ngày tải lên: 20/06/2014, 04:20

... reads genes and occurrenceare mapped Figure reads transfrags Fortheof gene are (N2) sequence nucleotides the compared transfrags GenesS4 showstiling and with UTR more Thiseach the file length gene ... Acknowledgements We thank Wolfgang Huber for critical comments and help with the data analysis, Sanger core sequencing for their sequencing efforts and the Fraser lab for comments and suggestions The authors ... Figure 3a for schematic and Figure 3b for an example) that is, triplets where exons A and C have NI of approximately and the middle exon B has an NI of

Ngày tải lên: 09/08/2014, 20:20

... chosen for the RT-nPCR Specificity of the multiplex RT-nPCR A fragment of 247 and 177 bp was amplified from CDV wild-type strain and the vaccine strain, respectively Two bands of 247 and 177 ... selected for amplification of the H gene of CDV by RTPCR with primers P5 (5'-CCAATTCATCCAAGCTGTCC-3') and P6 (5'-GGGATTTGAACGGTTACATGAG-3') The amplified products were cloned and sequenced, and the ... sensitivity[6,7] Primers P1 and P4 (specific for CDV and conserved among CDV species), primer P2 (specific for CDV wild-type strain), and primer P3, P5 (specific for CDV vaccine strain), were...

Ngày tải lên: 12/08/2014, 04:20

... http://www.virologyj.com/content/7/1/110 poxvirus gene expression Thus, alterations in gene expression and replication are likely to occur during CDV exposure, and, could result in mutations affecting ... intermediate and late replication complexes [29] and is thought to function as an early transcription factor docking platform [30,31] Vaccinia Early Transcription Factor (VETF), comprising D6R and A7L, ... pre-initiation complex formation and affect transcriptional regulation and promoter recruitment Capping methyltransferase The poxvirus mRNA capping machinery, encoded by the D1R and D12L genes in VACV,...

Ngày tải lên: 12/08/2014, 04:20