Nourseothricin sulfate is often used to select for cells with a mutation in the nat1 gene which confers resistance to Nourseothricin. It can be used in yeasts, bacteria, plant cells and filamentous fungi.. ...
Methods and compositions using populations of randomized modified FRT recombination sites to identify, isolate and/or characterize modified FRT recombination sites are provided. The recombinogenic mod
Self-resistance of the nourseothricin-producing strain Streptomyces noursei.: The nourseothricin producer Streptomyces noursei is resistant to its own antibioti
Chromosome segregation during meiosis is a complex process, which leads to production of four haploid gametes from two precursor cells. Reversible phosphorylation of proteins plays a crucial role in this process. The Schizosaccharomyces pombe Prp4 is an essential serine/threonine protein kinase, which belongs to the Clk/Sty family. To study the role of Prp4 in meiosis, we analysed chromosome segregation in a strain carrying conditional analog-sensitive allele of Prp4 protein kinase (prp4-as2). Our data show, that Prp4 protein kinase plays important role in chromosome segregation during meiosis, as revealed by enhanced missegregation of chromosomes in prp4-as2 mutant cells ...
KW: responding to my request for antibiotics useful for studies of genetics of resistance; Woodruff; Kane; streptomycin; streptothricin; grisein; jl 4/2/00 ...
This volume, which consists of 1,171 references on streptomycin for the years 1944 to 1948, 12 references dealing with actinomyces and 16 with streptothricin, a
Candida parapsilosis is a major cause of human disease, yet little is known about the pathogens virulence. We have developed an efficient gene deletion system for C. parapsilosis based on the repeated use of the dominant nourseothricin resistance marker (caSAT1) and its subsequent deletion by FLP-mediated, site-specific recombination. Using this technique, we deleted the lipase locus in the C. parapsilosis genome consisting of adjacent genes CpLIP1 and CpLIP2. Additionally we reconstructed the CpLIP2 gene, which restored lipase activity. Lipolytic activity was absent in the null mutants, whereas the WT, heterozygous, and reconstructed mutants showed similar lipase production. Biofilm formation was inhibited with lipase-negative mutants and their growth was significantly reduced in lipid-rich media. The knockout mutants were more efficiently ingested and killed by J774.16 and RAW 264.7 macrophage-like cells. Additionally, the lipase-negative mutants were significantly less virulent in infection ...
Glucosaminyl (N-acetyl) Transferase 1/GCNT1 Overexpression Lysate (Native). Tested Reactivity: Hu. Validated: WB. Backed by our 100% Guarantee.
Myc-DDK-tagged ORF clone of Homo sapiens glucosaminyl (N-acetyl) transferase 4, core 2 (GCNT4) as transfection-ready DNA - 10 µg - OriGene - cdna clones
InterPro provides functional analysis of proteins by classifying them into families and predicting domains and important sites. We combine protein signatures from a number of member databases into a single searchable resource, capitalising on their individual strengths to produce a powerful integrated database and diagnostic tool.
ConspectusResistance of pathogenic bacteria against currently marketed antibiotics is again increasing. To meet the societal need for effective cures, scientists are faced with the challenge of developing more potent but equally bacteria-specific drugs. Currently, most efforts are directed toward the modification of existing antibiotics, but ideally, compounds with a new mode of action are required. In this Account, we detail our findings in the area of novel metal-based antibiotics. Our strategy is based on the modification of simple antimicrobial peptides (AMPs) with organometallic agents, resulting in organometallic AMPs (OM-AMPs). Since bacteria have most likely never encountered these synthetically prepared unnatural organometallic agents, we anticipated that such agents could well become potentiating players in the antibiotics arena. Moreover, exploiting some of the particular properties of metal complexes should also help to elucidate the mode of action of small cationic AMPs, the ...
To investigate antibiotic resistance, the occurrence and distribution of class 1 and 2 integrons in multidrug- resistant Staphylococcus aureus isolates from hospitals in Tehran, Iran. The isolates were examined for susceptibility to antimicrobial agents. The mecA gene, class 1 and 2 integrons were detected by PCR. Integrase positive strains were further analysed for the presence of resistance gene cassettes using specific primers and were sequenced. Among 139 S.aureus isolates, 109 (78.4 %) and 112 (80.5 %) strains were considered as multidrug resistant and mecA positive, respectively. Class 1 integrons and internal variable regions were found in 72.6 % (101/139) and 97 % (98/101) and class 2 integrons and variable regions also in 35.2 % (49/139) and 65.3 % (32/49) of S.aureus clinical isolates, respectively. Twelve distinct cassette arrays were found, containing genes encoding resistance to β-lactams, aminoglycosides, streptothricin, trimethoprim, chloramphenicol,a putative glucose dehydrogenase
2-Oxocarboxylic acids, also called 2-oxo acids and alpha-keto acids, are the most elementary set of metabolites that includes pyruvate (2-oxopropanoate), 2-oxobutanoate, oxaloacetate (2-oxosuccinate) and 2-oxoglutarate. This diagram illustrates the architecture of chain extension and modification reaction modules for 2-oxocarboxylic acids. The chain extension module RM001 is a tricarboxylic pathway where acetyl-CoA derived carbon is used to extend the chain length by one. The chain modification modules RM002 (including RM032) and RM033, together with a reductive amination step (RC00006 or RC00036), generate basic and branched-chain amino acids, respectively. The modification module RM030 is used in the biosynthesis of glucosinolates, a class of plant secondary metabolites, for conversion to oxime followed by addition of thio-glucose moiety. Furthermore, the chain extension from 2-oxoadipate to 2-oxosuberate is followed by coenzyme B biosynthesis in methonogenic archaea ...
Phagocytosis is the process of taking in relatively large particles by a cell, and is a central mechanism in the tissue remodeling, inflammation, and defense against infectious agents. A phagosome is formed when the specific receptors on the phagocyte surface recognize ligands on the particle surface. After formation, nascent phagosomes progressively acquire digestive characteristics. This maturation of phagosomes involves regulated interaction with the other membrane organelles, including recycling endosomes, late endosomes and lysosomes. The fusion of phagosomes and lysosomes releases toxic products that kill most bacteria and degrade them into fragments. However, some bacteria have strategies to escape the bactericidal mechanisms associated with phagocytosis and survive within host phagocytes ...
My research interests are focused on liver metabolism and function as well as hepatic pathologies. We previously established an integrated 3D spheroid cell culture system for primary human hepatocytes (PHH) in which cells remain viable and functionally stable for multiple weeks. Importantly, we recently showed that PHH in this model exhibited superior sensitivity to hepatotoxic agents compared to other emerging cell models, such as HepG2 and HepaRG cells, and were faithfully reproducing in vivo drug toxicity mechanisms in man. Furthermore, using a combination of untargeted and targeted metabolomics, we showed that the endogenous and xenobiotic metabolic signatures of PHH were maintained in 3D spheroids, thus allowing to comprehensively study drug-induced molecular effects on cellular metabolism and to investigate mechanisms of drug action. The results indicate that the 3D PHH spheroid system faithfully mimics heptic phenotypes in vivo and can be utilized for long-term analyses of drug ...
Treatment of bacterial infections with antibiotics is universally accepted as one of (if not THE) most significant contributions of medical intervention to reducing mortality and morbidity during last century. Surprisingly, basic knowledge about how antibiotics kill or prevent the growth of bacteria is only just beginning to emerge and the dose and term of antibiotic treatment has long been determined by clinicians empirically and intuitively. There is a recent drive to theoretically and experimentally rationalize antibiotic treatment protocols with the aim to them and to design protocols which maximize antibiotics efficacy while preventing resistance emergence. Central to these endeavors are the pharmacodynamics of the antibiotic(s) and bacteria, PD (the relationship between the concentration of the antibiotic and the rate of growth/death of bacteria), and the pharmacokinetics of the antibiotic, PK (the distribution and change in concentration of the antibiotics in a treated host) of each ...
Infection]] with one of the streptococcal [[bacteria]]. Strep infection can cause [[pus]]-filled abscesses on the [[skin]] or internal organs, and can migrate through the [[blood]] to infect the [[heart]], meninges, and other areas. Treatment is with [[antibiotic]]s and drainage of [[abscess]]es as needed ...
Aerobic and anaerobic cultivation techniques, 16S rDNA-based phylogeny, and fluorescent in situ hybridization were used to describe the phylogenetic diversity and physiological versatility of lotic microbial aggregates (river snow) obtained from the river Elbe. In the course of the year the river snow community changed. It was characterized by a great bacterial diversity in spring, the predominant occurrence of algae in summer and reduction of the total bacterial cell count in autumn and winter. In all river snow samples, more than 70% of the bacteria counted with the general DNA stain DAPI also hybridized with the Bacteria-specific probe EUB338. In situ analysis of the bacterial river snow community with a comprehensive suite of specific rRNA-targeted probes revealed population dynamics to be governed by seasonal factors. During all seasons, beta-Proteobacteria constituted the numerically most important bacterial group forming up to 54% of the total cell counts. In contrast to this, the ...
5. Oh H, Ozkirimli E, Shah K, Harrison ML, Geahlen RL. Generation of an analog-sensitive Syk tyrosine kinase for the study of signaling dynamics from the B cell antigen receptor. J Biol Chem. 2007 Nov 16;282(46):33760-8. Epub 2007 Oct 3. PubMed PMID: 17913708 ...
Puromycin resistance vector confers puromycin resistance; can be used as a dominant selectable marker to select for stably transformed mammalian cells.
Puromycin resistance vector confers puromycin resistance; can be used as a dominant selectable marker to select for stably transformed mammalian cells.
From Irina Borodinas lab, the EasyClone v2 kit is an expanded toolkit of integrative vectors for stable gene expression in yeast.
Most of the Moosers here on Bargainmoose are familiar with Well.ca, the online store from which you can purchase a plethora of items; everything from su...
There is one other thing i have just noticed that may or may not cause a problem. The button i have picked for the Switch has 3 states in ableton On/Held/Off they relate only if there is a Track in ableton for the column on the APC, the button i have picked is on Track 7 and so with no Ableton Track in the project it only sends a note on or off, the codes in midi capture look like this for Track 1 90 30 7F (on), 90 30 00 (held) 80 30 7F (off), however on Track 7 i only get 96 30 7F (on), 86 30 7F (off). i have tried to manually add to 00 which is the obvious velocity but it doesnt work as there is only a note on/off message being sent. How can i recreate this function so that MT can see it without an Ableton Track loaded ...
First-time academic entrepreneurs face a circular problem: while theyre continually sought after by VCs and pharmas seeking the next
studied using thirteen microsatellite markers and results compared to six European R. secalis populations. The genetic structure of P. teres was examined with four AFLP markers and compared to populations from Finland and Russia. Both studies revealed a significant differentiation between the Icelandic and European populations. The mean Fst value between the Icelandic R. secalis population and five Scandinavian populations was 0.165 and pairwise differentiation between the P. teres populations from Iceland and Finland was 0.640. Of the P. teres populations analyzed the gene diversity was highest in Iceland, with a value of 0.26, compared to 0.17 and 0.04 in the Russian and ...
Cdc7-Dbf4 is an essential protein kinase complex required for every single origin firing. As a target of the intra-S checkpoint, Cdc7 kinase activity has also been implicated in the response to replication fork stress, with a role in translesion DNA synthesis (TLS). We have examined the role of Cdc7 in the regulation of replication forks, particularly in response to MMS, which normally stalls replication forks and inhibits late origin firing. We find that replication forks proceed as fast as with no damage along an MMS-damaged template both in cdc7as3 and cdc7-1/mcm5-bob1 cells. However the DNA synthesis in cdc7-1/mcm5-bob1 in MMS is defective, indicated by the slower recovery after MMS by PFGE, suggesting the replication is incomplete. These deregulated forks did not rely on TLS pathway but are dependent on both helicase and E3 ligase function of Rad5 for continued fork progression along MMS-damaged DNA, demonstrating a role for Rad5 at the replication fork. Phosphorylation of MCM2 by DDK was ...
General Information 1. What is growth hormone? Growth hormone (GH) is a single polypeptide hormone secreted from the cells of the pituitary gland. GHs in human are present as isomers of 27, 22, 20, 17, and 5 kDa. GH of 22 kDa, called somatropin, is the major component of GHs produced by human pituitary and used for the therapy. This kind of GH is composed of 191 amino acids and contains four -helices arranged in a left- handed bundle orientation. Two disulfide bridges are located at Cys53-Cys165 and Cys182-Cys189 in GH, and the Cys residues make up the loop structure of the GH molecule. The main effect of GH is to promote postnatal longitudinal growth. Hyposecretion of GH can lead to dwarfism. The growth-promoting effects of GH result from GHs diverse and pleiotropic effects on cellular metabolism and differentiation. GH is known to regulate the lipid, carbohydrate, nitrogen, and mineral metabolism within a cell. Many of GH actions are mediated by the activation of insulin-like growth
Step 1 - Thawing Cells: Thaw competent cells on ice. Step 2 - Adding cells: Add 50 µL of thawed competent cells into pre-chilled 2ml tube. Step 3 - Addition of BioBrick: Add 1 - 2 µL of the resuspended DNA to the 2ml tube. Pipette up and down a few times, gently. Make sure to keep the competent cells on ice. Step 4 - Incubation: Close tube and incubate the cells on ice for 30 minutes. Step 5 - Heat Shock: Heat shock the cells by immersion in a pre-heated water bath at 42ºC for 60 seconds. Step 6 - Incubation: Incubate the cells on ice for 5 minutes. Step 7 - Add media: Add 200 μl of SOC media or LB broth Step 8 - Incubation: Incubate the cells at 37ºC for 2 hours while the tubes are rotating or shaking. Step 9 - Label plates: Label two petri dishes with LB agar and the appropriate antibiotic(s) with the part number, plasmid backbone, and antibiotic resistance. Plate 20 µl and 200 µl of the transformation onto the dishes, and spread. This helps ensure that you will be able to pick out a ...
Crown-gall tumor initiation by Agrobacterium tumefaciens is inhibited by cell walls from normal dicotyledonous plants but not by cell walls from crown-gall tumors apparently because of bacterial adherence or nonadherence, respectively, to the different cell walls. Cell walls from normal and tumor tissues in culture also show this difference, indicating that the two types of tissue stably maintain this difference under these conditions. Habituated tissue cultures, which resemble crown-gall tumor cultures, however, form cell walls that are inhibitory like those of the normal cultures from which they are derived. Monocotyledonous plants do not act as hosts for Agrobacterium and bacteria-specific inhibition is not shown by cell walls from several species of grass, a monocot family. Cell wallsfrom "embryonic" tissues (dicot seedlings less than 2 centimeters long), unlike those from older seedlings, are non-inhibitory. Crown-gall tumors thus resemble embryonic tissues in this respect.. ...
Large-scale production and purification of recombinant proteins by cell cultures represent a key-area in manufacturing field. The production process still has several drawbacks affecting cost/efficiency. In this work, three modular systems were designed to overcome some of these bottlenecks. A library of self-inducible promoters was built and characterized to start the peptide production at a desired culture density, without expensive inducer molecules. Two standard integrative vectors were realized to insert BioBrick parts in user-defined positions of E. coli or S. cerevisiae genome, to ensure genetic stability without using selection markers. Finally, two promising techniques were combined for an "in-cell" protein purification: PolyHydroxyAlkanoate (PHA) granules were used as a substrate for PHA-binding peptides (Phasins) fused to the target protein, thus replacing affinity resins/columns and tags, while a pH-based self-cleaving peptide (Intein) was used instead of a protease cleavage site. ...
Large-scale production and purification of recombinant proteins by cell cultures represent a key-area in manufacturing field. The production process still has several drawbacks affecting cost/efficiency. In this work, three modular systems were designed to overcome some of these bottlenecks. A library of self-inducible promoters was built and characterized to start the peptide production at a desired culture density, without expensive inducer molecules. Two standard integrative vectors were realized to insert BioBrick parts in user-defined positions of E. coli or S. cerevisiae genome, to ensure genetic stability without using selection markers. Finally, two promising techniques were combined for an in-cell protein purification: PolyHydroxyAlkanoate (PHA) granules were used as a substrate for PHA-binding peptides (Phasins) fused to the target protein, thus replacing affinity resins/columns and tags, while a pH-based self-cleaving peptide (Intein) was used instead of a protease cleavage site. ...
Nepalgunj has a sub-tropical climate. Temperatures sometimes exceed 40°C (104°F) from April to June. During the rainy season-arriving in June and lasting into September-it is less hot but sometimes very humid. Winter is usually pleasant while the sun is out. It sometimes is foggy and overcast; then it can be chilly with temperatures below 10°C (41°F) but no frost. The highest temperature ever recorded in Nepalgunj was 45.0 °C (113.0 °F) on 16 June 1995, while the lowest temperature ever recorded was −0.3 °C (31.5 °F) on 9 January 2013.[4] ...
A classical biological control project was undertaken against the filbert aphid, Myzocallis coryli (Goetze), in western Oregon. Hazelnut orchards in Spain, France, and Italy were searched for natural enemies of the aphid, and a parasitic wasp, Trioxys pallidus Haliday, was imported, quarantined, mass-reared, and released in Oregon orchards. The wasp successfully attacks and completes its development on the Oregon race of the filbert aphid. A greenhouse culture of the parasitoid has been maintained continuously for over 50 generations, and approximately 30,000 adult wasps have been released in the Willamette Valley. Overwintering survival has been documented in at least eight different locations. In three commercial orchards, the parasitoids reduced aphid populations by 26-32%. The newly imported biotype of T.pallidus attacking the filbert aphid was compared with the previously established biotype attacking the walnut aphid. The two races are morphologically indistinguishable, and were shown to ...
The antifungal polyene macrolide nystatin is produced by Streptomyces noursei ATCC 11455. The nystatin biosynthesis gene cluster of Streptomyces noursei has been cloned and sequenced, and a biosynthesis route has been predicted. In the present work, investigation of genes presumably involved in post-PKS modifications of nystatin is described. The aim of this work was to better understand the nystatin biosynthesis and to further use this information for generation of novel nystatin analogues. Two PKS-modifications of the nystatin molecule were targeted in this study: glycosylation with mycosamine at C-19 and oxidation of the exocyclic methyl group at C-16.. Two genes putatively involved in mycosamine biosynthesis (NysDIII and NysDII) and one in attachment of mycosamine to the nystatin aglycone (nysDI) have been identified in the nystatin gene cluster. Their functions have been suggested, respectively, as a putative mannose dehydratase, aminotransferase and a glycosyltransferase. The deoxysugar ...
Here, we performed high-throughput drug-screening to identify new non-toxic mitochondrial inhibitors. This screening platform was specifically designed to detect compounds that selectively deplete cellular ATP levels, but have little or no toxic side effects on cell viability. Using this approach, we identified DPI (Diphenyleneiodonium chloride) as a new potential therapeutic agent. Mechanistically, DPI potently blocks mitochondrial respiration by inhibiting flavin-containing enzymes (FMN and FAD-dependent), which form part of Complex I and II. Interestingly, DPI induced a chemo-quiescence phenotype that potently inhibited the propagation of CSCs, with an IC-50 of 3.2 nano-molar. Virtually identical results were obtained using CSC markers, such as CD44 and CD24. We further validated the effects of DPI on cellular metabolism. At 10 nM, DPI inhibited oxidative mitochondrial metabolism (OXPHOS), reducing mitochondrial driven ATP production by >90%. This resulted in a purely glycolytic phenotype, with
There are four active projects in my lab, all devoted to taking advantage of the ecological and behavioral diversity of the microbial world. One is studying interactions between gut microbes and their hosts, using a new model system, planaria (flatworms) as the experimental animals. The second is a bioremediation project, isolating bacteria which catabolize gasoline as their source of carbon and energy, and using those bacteria alone or in combination to render gasoline contaminated soil safe for normal use. The third project uses bacteriophage (bacteria-specific viruses) as an antibacterial agent for the skin disease acne, targeting the bacterial species that causes serious acne lesions. The final project is working with bacteriophage specific for food-borne pathogens, developing kitchen-based protocols to decontaminate produce like lettuce or alfalfa sprouts which are not cooked before consuming. ...
Sengupta S and Chandra T.S , Molecular and structural characterization of GTP-cyclohydrolase II in Eremothecium ashbyi NRRL Y-1363: cDNA cloning, comparative sequence analysis and molecular modeling . Fungal Biology 2010 doi:10.1016/j.funbio.2010.06.006 Copyright © 2010 The British Mycological Society Published by Elsevier Ltd (originally called Mycological Research ...
... The 30-Minute Tip for Produce My Papers Even should you prepare a report to find an F on it, you can expect to
Yep, full Oehler, aka professional instruments. The middle finger position is indeed sharp for Austrian models. But for German models, the level of sharpness is not severer than the lowness of low F on Boehm systems (at least for my clarinets), which most people accept for so many years until recently manufacturers begin to add the correction key. Also, I would like to mention a not widely known theory here. As you all know, clarinet is the only transposing instrument in the woodwind section. For all other members of this section (i.e, flute, oboe and bassoon), without any doubt, their key systems are designed, ideally, to play the C major scale most easily. By realizing this, it is not hard to understand why F sharps can be played easily on Oehler system while F naturals are not. This can be also observed for the third space C sharp (and the high C sharp). On Boehm, the transition from middle B to C sharp is not very smooth and in fact the trill of B/C sharp is in theory impossible because it ...
Yep, full Oehler, aka professional instruments. The middle finger position is indeed sharp for Austrian models. But for German models, the level of sharpness is not severer than the lowness of low F on Boehm systems (at least for my clarinets), which most people accept for so many years until recently manufacturers begin to add the correction key. Also, I would like to mention a not widely known theory here. As you all know, clarinet is the only transposing instrument in the woodwind section. For all other members of this section (i.e, flute, oboe and bassoon), without any doubt, their key systems are designed, ideally, to play the C major scale most easily. By realizing this, it is not hard to understand why F sharps can be played easily on Oehler system while F naturals are not. This can be also observed for the third space C sharp (and the high C sharp). On Boehm, the transition from middle B to C sharp is not very smooth and in fact the trill of B/C sharp is in theory impossible because it ...
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