core rna polymerase lacking d e and purified d protein

... Espo, Finland) Proteins were detected using chemiluminescence reagents (Pierce) Antibodies GIA, E2 4C and H3B, or N20 were used for the detection of Pol e and RNA pol II, respectively For dephosphorylation, ... complexes were extracted with phenol, precipitated with acetone, collected at °C and washed three times The dried protein nucleic acid pellet was resuspended by boiling in denaturation buffer (50 ... PAGE, transferred to poly(vinylidene difluoride) membrane and exposed to BioMax MS film (Kodak, Vantaa, Finland) After autoradiography, proteins were detected using enhanced chemiluminescence reagents...

Ngày tải lên: 07/03/2014, 11:20

... Sies and Lester Packer Volume 348 Protein Sensors and Reactive Oxygen Species (Part B: Thiol Enzymes and Proteins) Edited by Helmut Sies and Lester Packer Volume 349 Superoxide Dismutase Edited ... Sidney Fleischer and Lester Packer Volume LVI Biomembranes (Part G: Bioenergetics) Edited by Sidney Fleischer and Lester Packer Volume LVII Bioluminescence and Chemiluminescence Edited by Marlene ... also be monitored with RNAse, because downstream movement of the polymerase relative to the RNA exposes more RNA to digestion Experiments are done as described for the exonuclease digestions: ECs...

Ngày tải lên: 11/04/2014, 10:30

... prior to making measurements The measurements were made in the presence and absence of urea Each experiment was performed at least three times with different protein preparations Size-exclusion chromatography ... urea is blue shifted because the tryptophan residues are buried in the hydrophobic interior of the protein As the protein unfolds, the tryptophan residues become more exposed to the solvent and ... urea, the dashed line corresponds to the samples without treatment, and the dotted line represents the samples treated with high pressure and returned to atmospheric pressure The flow rate was 0.3...

Ngày tải lên: 19/02/2014, 12:20

... Studies with CatE-deficient and CatD-deficient mice have provided additional evidence of the association of these enzymes with different physiological effects CatD-deficient mice develop massive intestinal ... CatE mRNA in different cell lines To determine the expression of CatE at the mRNA level in different cell lines, RT-PCR was performed using RNA extracted from DCs (monocyte-derived human dendritic ... was digested by CatE, CatD or subcellular fractions (as described in Experimental procedures) The peptide fragments thus generated were separated by RP-HPLC using fluorescence detection (kex ¼...

Ngày tải lên: 07/03/2014, 09:20

... unlabeled methionine for different periods of time up to 24 h At the end of the chase, cells and culture media were collected separately The cell lysate and culture medium were subjected to immunoprecipitation ... and the ER-retention sequence (DER-ret.) (C) were pulse-labeled with [35S]methionine ⁄ cysteine for 30 and chased for the times indicated The cell lysate and the culture medium were immunoprecipitated ... wild-type cathepsin E were pulse-labeled with [35S]methionine ⁄ cysteine for 30 and chased with nonradiolabeled medium for the times indicated The cell lysate and culture medium were immunoprecipitated...

Ngày tải lên: 16/03/2014, 14:20

... the large number of characterized CTDkinases, few CTD-phosphatases have been identified (Table and references therein); most studies concern the TFIIF-dependent CTD phosphatase (FCP1) [59] Recently, ... phosphorylate both serine and serine [41,44] Efficiencies of the different kinases are influenced by the position of the targeted repeats within the CTD [38,45] and by the neighboring residue’s identity ... Slightly different conclusions were reached when determining phosphoepitopes that were generated on purified RNAP II For example, CDK7 (TFIIH) generates H14 epitope (serine 5) as predicted [46,47]...

Ngày tải lên: 17/03/2014, 10:20

... for 36–48 h Whole cell extracts were prepared as described in Materials and methods and 90 lg were loaded per lane Proteins were separated on 10% SDS/PAGE gels and analyzed by Western blotting with ... (W) and eluate (E) fractions were analyzed by Western blotting with the indicated antibodies The figure shows one of three independent experiments (lines a d) or one of two independent experiments ... expressed in 293 cells and whole cell extracts were prepared and 110 lg were loaded per lane Proteins were resolved on 10% SDS/PAGE gels and analyzed by Western blotting with anti-FLAG (lanes 1–4) and...

Ngày tải lên: 17/03/2014, 10:20

... DNA strands were separated by size on 10% denaturing PAGE, the nonbiotinilated strand was eluted and used for the next SELEX round In the Rifdirected SELEX experiment each round of the selection ... nucleotide deletion at the same site The sequence motif identical in aptamers E9 and E1 2 is underlined treated with phenol and chloroform and ethanol precipitated The resulting enriched library ... that formed stable complexes with RNAP RNAP–DNA complexes were eluted with imidazole, recovered oligonucleotides were amplified by PCR and used in the next round of selection To avoid selection...

Ngày tải lên: 23/03/2014, 13:20

... if added when the enzyme is already bound to DNA [16] As shown in Fig 6, all three compounds totally inhibited RNA synthesis when added before DNA, whereas Fig E ect of GE23077 and other RNA polymerase ... controls; d, compounds were added before the reaction start, marked by the addition of DNA to mixtures containing all the other components and the indicated inhibitor; m, nucleotides and the indicated ... Rifampicin and streptolydigin were used as reference inhibitors of, respectively, transcription initiation and chain elongation As expected, all three compounds behaved similarly when added to the reaction...

Ngày tải lên: 30/03/2014, 15:20

... acid The substrate analogues, presumed products, and phosphate ion were eluted at the beginning of the run and collected by hand The mixture was dried by evaporation under vacuum and redissolved ... of the wild-type Protein determination and enzyme assay Protein concentrations were determined by the method of Bradford using bovine serum albumin as standard [6] The Ó FEBS 2004 D- Ornithine assay ... gel-purified and assembled in a second round of PCR using oligonucleotides 40 and 41 as primers and cotemplates The PCR product was purified, restricted with SpeI and BamHI, and ligated with SpeI/BamHI-restricted...

Ngày tải lên: 30/03/2014, 15:20

... Edited by DAVID BARNES AND DAVID A SIRBASKU VOLUME 148 Plant Cell Membranes Edited by LESTER PACKER AND ROLAND DOUCE VOLUME 149 Drug and Enzyme Targeting (Part B) Edited by RALPH GREEN AND KENNETH ... SIDNEY FLZISCHERAND LESTER PACKER VOLUME LVI Biomembranes (Part G: Bioenergetics) Edited by SIDNEY FLEISCHER AND LESTER PACKER VOLUME LVII Bioluminescence and Chemiluminescence Edited by MARLENE ... overlapped the promoters they regulated led to a straightforward model for repression: the presence of repressor should sterically prevent the binding of RNAP Detailed kinetic evidence that repressors...

Ngày tải lên: 11/04/2014, 10:30

... and C) Edited by DONALD B McCORMICK AND LEMUEL D WRIGHT VOLUME XIX Proteolytic Enzymes Edited by GERTRUDE E PERLMANNAND LASZLO LORAND VOLUME XX Nucleic Acids and Protein Synthesis (Part C) Edited ... Prostaglandins and Arachidonate Metabolites Edited by WILLIAM E M LANDS AND WILLIAM L SMITH VOLUME87 Enzyme Kinetics and Mechanism (Part C: Intermediates, Stereochemistry, and Rate Studies) Edited ... F: Bioenergetics) Edited by SIDNEY FLEISCHER AND LESTER PACKER xxii METHODS IN ENZYMOLOGY VOLUME LVI Biomembranes (Part G: Bioenergetics) Edited by SIDNEY FLEISCHER AND LESTER PACKER VOLUME LVII...

Ngày tải lên: 11/04/2014, 10:30

... Sies and Lester Packer Volume 348 Protein Sensors and Reactive Oxygen Species (Part B: Thiol Enzymes and Proteins) Edited by Helmut Sies and Lester Packer Volume 349 Superoxide Dismutase Edited ... Protein and Nucleic Acid Sequences Edited by Russell F Doolittle Volume 184 Avidin-Biotin Technology Edited by Meir Wilchek and Edward A Bayer Volume 185 Gene Expression Technology Edited by David ... by Bernardo Rudy and Linda E Iverson Volume 208 Protein DNA Interactions Edited by Robert T Sauer Volume 209 Phospholipid Biosynthesis Edited by Edward A Dennis and Dennis E Vance methods in enzymology...

Ngày tải lên: 11/04/2014, 10:30

... primer extension The first base of the predominate RNA transcript of the indicated species is labeled +1 Conserved residues are indicated blue, red and green lettering For all the indicated species, ... ladder size marker Using the marker as a guide, the kb region of the EcoR I digested MDCK DNA lane was excised from the gel followed by recovery of the DNA from the gel sample The recovered DNA ... then were analyzed on agarose gels to identify colonies which produced 500 bp products, the size predicted from the 18S rRNA gene sequence One such clone, designated pK9PolI, was determined by...

Ngày tải lên: 18/06/2014, 18:20

... primer extension The first base of the predominate RNA transcript of the indicated species is labeled +1 Conserved residues are indicated blue, red and green lettering For all the indicated species, ... ladder size marker Using the marker as a guide, the kb region of the EcoR I digested MDCK DNA lane was excised from the gel followed by recovery of the DNA from the gel sample The recovered DNA ... then were analyzed on agarose gels to identify colonies which produced 500 bp products, the size predicted from the 18S rRNA gene sequence One such clone, designated pK9PolI, was determined by...

Ngày tải lên: 20/06/2014, 01:20

... produced by Dicer [19] and so they are likely to be produced by a Dicer-independent mechanism Different members of the PIWI protein family show a preference for binding to either the sense or ... thus strengthen­ ng the argument that Elp1 provides an RNAi dependent RNA polymerase activity that generates the antisense strand Unlike the gene for canonical RdRP, the elp1 gene is conserved in ... convert mRNA into dsRNAs that are degraded to generate siRNAs Cell 2001, 107:297-307 10 Lipardi C, Paterson BM: Identification of an RNA- dependent RNA polymerase in Drosophila involved in RNAi and...

Ngày tải lên: 09/08/2014, 20:20

... histone H3 (b), H3K36me3 (c), and FAIRE (d) were used to generate average spliced gene profiles that were grouped into ten ranked bins based on Affymetrix expression data Scales for the relative data ... range from each expression bin were used to generate the plots Identical data plotted on the same absolute y-scale for all expression bins is presented for average spliced and unspliced genes ... unspliced genes (2,218 and 2,598 genes, respectively) Lists of spliced/unspliced genes were then ranked in descending order and split into 10 equal groups (approximately 220 and 260 genes per group...

Ngày tải lên: 09/08/2014, 23:20

... Nurse, Cancer Research UK Your research papers will be: available free of charge to the entire biomedical community peer reviewed and published immediately upon acceptance cited in PubMed and archived ... AA-Carried out the data collection procedures FG-Carried out the data collection procedures GC-Supervised the data collection procedures and conducted bibliographic research All authors read and approved ... parameter, with the confidence interval (C.I.) at 95%, Amplitude (A), expressed in the same unit as the relative Mesor, and Acrophase (φ), obtained with the single Cosinor method [12] and expressed...

Ngày tải lên: 10/08/2014, 09:20

... to expand the available ARV sequence database, determine sequences of the ARV RdRp protein, and to delineate conserved structure/ function features of this key viral-encoded enzyme Methods Cells ... centrifuged immediately at 10,000 × g for 30 minutes The supernatants were removed and pellets were dried and dissolved in ddH2O for cDNA synthesis Full-length cDNA copies of each L2 genome segment were ... reactions were designed from newly obtained sequences to completely sequence each full-length PCR product in both directions Sequences nearer the ends of each segment were determined from PCR products...

Ngày tải lên: 12/08/2014, 04:21

... positive ANA (speckled and nucleolar pattern) A year later she developed proximal scleroderma and visited the UFCAD RP developed six months later Case 3: A 57-year-old female, who had a 10-year ... fibrillarin, and -Th/To, have been reported to be associated with scleroderma (systemic sclerosis, SSc); some are considered highly specific disease markers while others are considered relatively specific ... were positive but no scleroderma or RP was noted Prednisolone (40 mg/day) was started and her condition was followed at her local clinic She developed proximal scleroderma and scleroderma renal...

Ngày tải lên: 12/08/2014, 17:22