Journal Article Deletion screening of the Duchenne muscular dystrophy locus via multiplex DNA amplification Get access Jeffrey S. Chamberlain, Chamberlain 1Institute for Molecular Genetics, Baylor College MedicineHouston, TX 77030, USA Search other works by this author on: Oxford Academic PubMed Google Scholar Richard A. Gibbs, Gibbs Joel E. Rainer, Rainer Phi Nga Nguyen, Nguyen USA2Institute Howard Hughes Medical institute, C. Thomas Nucleic Acids Research, Volume 16, Issue 23, 9 December...

Conformational control of biomolecular activities can reveal functional insights and enable the engineering novel activities. Here we show that conformational through intramolecular cross-linking a helicase monomer with undetectable unwinding activity converts it into superhelicase unwind thousands base pairs processively, even against large opposing force. A natural partner enhances is shown to achieve its stimulating role also by selectively stabilizing active conformation. Our work...

Several staphylococcal plasmids from different incompatibility (inc) groups which replicate by a rolling circle mechanism each specify replication initiator protein (Rep) is homologous with that of the inc3 tetracycline resistance plasmid pT181. The rep gene sequences six pT181-like are known, encoding proteins molecular mass 38 kDa 62% overall amino acid sequence identity. initiation in vivo Rep specific, acting trans only at cognate origin (ori) plasmid. Previous studies vitro RepC pT181...

The plasmid replication initiator protein, RepD, greatly stimulates the ability of DNA helicase, PcrA, to unwind lengths DNA. Unwinding begins at oriD, double-stranded origin that RepD recognizes and covalently binds initiate replication. Using a combination plasmids containing oriD oligonucleotide structures mimic parts kinetics nicking separation have been determined, along with coupling ratio between base ATP hydrolysis. At 30 degrees C, rate is 1.0 s(-1), translocation approximately bp...

The first synthesis of dolabelide C (1), a cytotoxic marine macrolide, is reported utilizing phosphate tether-mediated approach. Bicyclic phosphates (S,S,SP)-5 and (R,R,RP)-5 serve as the central building blocks for construction two major 1,3-anti-diol subunits in 1 through selective cleavage pathways, regioselective olefin reduction, cross-metathesis. Overall, phosphate-mediated processes provided copious amounts both allowing detailed RCM macrocyclization study to 24-membered macrolactone 1.

ABSTRACT Mobilization of the staphylococcal plasmid pC221 requires at least one plasmid-encoded protein, MobA, in order to form a relaxosome. and closely related plasmids also possess an overlapping reading frame encoding protein 15 kDa, termed MobC. By completing nucleotide sequence pC223, we have found further example this small gene knockouts shown that MobC is essential for relaxosome formation mobilization both pC223. Primer extension analysis has been used identify nic site these...

The complete nucleotide sequence of a naturally occurring Staphylococcus aureus plasmid, pT48 (from S. strain T48), has been determined. 2475 bp plasmid confers inducible resistance to macrolide-lincosamide-streptogramin B (MLS) type antibiotics. It is similar the constitutive MLS pNE131, from epidermidis and shows homology with plasmids pSN2 pE194. contains palA structure homologous that on pT181. open reading frame, ORF B, within region frameshifted C-terminus, relative resulting in...

Antibiotic resistance in pathogenic bacteria is a continual threat to human health, often residing extrachromosomal plasmid DNA. Plasmids of the pT181 family are widespread and confer various antibiotic resistances Staphylococcus aureus. They replicate via rolling circle mechanism that requires multi-functional, plasmid-encoded replication protein initiate replication, recruit helicase site initiation terminate after DNA synthesis complete. We present first atomic resolution structures three...

Summary The nicking of the origin transfer ( oriT ) is an essential initial step in conjugative mobilization plasmid DNA. In case staphylococcal pC221, by plasmid‐specific MobA relaxase facilitated DNA‐binding accessory protein MobC; however, role MobC this process currently unknown. study, site binding was determined DNase I footprinting. interacts with DNA at two directly repeated 9 bp sequences, mcb1 and mcb2 , upstream nic site, additionally a third, degenerate repeat within mobC gene,...

ABSTRACT The isolation of plasmid-protein relaxation complexes from bacteria is indicative the plasmid nicking-closing equilibrium in vivo that serves to ready plasmids for conjugal transfer. In pC221 and pC223, components required site- strand-specific nicking at oriT are MobC MobA. order investigate minimal requirements absence host-encoded factors, reactions were reconstituted vitro. Purified MobA MobC, presence Mg 2+ or Mn , found nick with a concomitant phosphorylation-resistant...

Construction of the C15−C30 subunit dolabelide utilizing a temporary phosphate tether is described. Two routes are reported that make use orthogonal protecting- and leaving-group properties innate to esters. One route relies on selective terminal oxidation, while second utilizes CM/selective hydrogenation sequence. Both depend highly regio- diastereoselective cuprate addition set requisite stereochemistry at C22.

Relative biodistribution of FDA-approved innovator and generic sodium ferric gluconate (SFG) drug products was investigated to identify differences in tissue distribution iron after intravenous dosing rats. Three equal cohorts 42 male Sprague-Dawley rats were created with each cohort receiving one three treatments: (1) the SFG product dosed intravenously at a concentration 40 mg/kg; (2) (3) saline equivalent volume products. Sampling time points 15 min, 1 h, 8 week, two weeks, four six weeks...

The study of plasmid DNA replication not only has yielded basic information on the mechanisms involved in maintenance these elements bacteria, but also led to development several model systems that have provided insights into biochemical processes and genetic regulation general. various studied date show replicons vary considerably their modes replication, at least with respect copy number directionality (Helinski 1976). A clear requirement each case, however, is a mechanism ensures...

Conference Article| October 01 1988 Identification of the tyrosine residue involved in bond formation between replication origin and initiator protein plasmid pC221 CHRISTOPHER D. THOMAS; THOMAS 1Department Biochemistry, University Leicester, Road, Leicester LE1 7RH, U.K. Search for other works by this author on: This Site PubMed Google Scholar DEBORAH F. BALSON; BALSON W. V. SHAW * *To whom correspondence should be addressed. Author article information Publisher: Portland Press Ltd...

Plasmid encoded replication initiation (Rep) proteins recruit host helicases to plasmid origins. Previously, we showed that RepD recruits directionally the PcrA helicase pC221 oriD, remains associated with it, and increases its processivity during unwinding. Here show forms a complex extending upstream downstream of core oriD. Binding causes remodelling region from oriD forming ‘landing pad' for PcrA. is recruited by this extended RepD–DNA via an interaction at site. appears have weak...

A rapid, sensitive and specific ultrafiltration inductively-coupled plasma mass spectrometry (UF-ICP-MSICP-MS) method was developed validated for the quantification of non-transferrin bound iron (NTBI), transferrin (TBI), drug (DI) total (TI) in same rat serum sample after intravenous (IV) administration gluconate nanoparticles sucrose solution (Ferrlecit®). Ultrafiltration with a 30 kDa molecular cut-off filter used cleanup. Different elution solvents were to separate each form from serum....

A series of homoleptic tris-chelate complexes ML3 (M = Al or rare earth; L chiral achiral oxazolyl phenolate -naphtholate) is reported. In all cases, crystallize as mer-isomers and complete diastereoselectivity observed on crystallization the complexes: with Λ-helicity at metal where (S)-oxazolylphenolate. Complexes have been characterized in solution by NMR spectroscopy, demonstrating rapid ligand exchange ambient temperature for earth complexes, slow timescale Al; cases mer-isomer...

A sensitive, specific and rapid ultra-performance liquid chromatography tandem mass spectrometry method has been developed validated for the determination of dexamethasone (DEX) in ocular tissues rabbits.

Conference Article| October 01 1988 Expression and purification of the rat liver ferritin light chain after cloning in Escherichia coli C. D. THOMAS; THOMAS *Department Biochemistry, University Leicester, Road, Leicester LE1 7RH, U.K. Search for other works by this author on: This Site PubMed Google Scholar W. V. SHAW; SHAW M. LAWSON; LAWSON ‡Department The University, Sheffield S10 2TN, A TREFFRY; TREFFRY P. J. ARTYMIUK; ARTYMIUK HARRISON § §To whom correspondence should be addressed....

DNA topoisomerase IV removes undesirable topological features from molecules in order to help maintain chromosome stability. Two constructs of 56 and 59 kDa spanning the DNA-cleavage domain A subunit Staphylococcus aureus (termed GrlA56 GrlA59) have been crystallized. Crystals were grown at 291 K using sitting-drop vapour-diffusion technique with PEG 3350 as a precipitant. Preliminary X-­ray analysis revealed that crystals belong space group P21, diffract resolution 2.9 Å possess unit-cell...