Search results for "Prime"
showing 10 items of 853 documents
Production and characterization of the recombinant Sphingomonas chlorophenolica pentachlorophenol 4-monooxygenase.
2001
Abstract Pentachlorophenol 4-monooxygenase (PCP4MO) from Sphingomonas chlorophenolica is a flavoprotein that hydroxylates PCP in the presence of NADPH and oxygen. In order to investigate the structure and function of active site, recombinant PCP4MO (rePCP4MO) was produced in Escherichia coli as a glutathione S-transferase (GST) fusion protein. Moreover, a tobacco etch virus (TEV) protease cleavage site (EKLYFQG) was introduced into GST-PCP4MO and a his-tagged TEV protease was employed. Hence, a two-step purification protocol was developed which allowed obtaining 15–20 mg of rePCP4MO from 1 L culture. The rePCP4MO revealed identity with native enzyme by SDS–PAGE and N-terminal sequence analy…
The E3 Ubiquitin Ligase MID1 Catalyzes Ubiquitination and Cleavage of Fu
2014
Sonic Hedgehog (SHH)-GLI signalling plays an important role during embryogenesis and in tumorigenesis. The survival and growth of several types of cancer depend on autonomously activated SHH-GLI signalling. A protein complex containing the ubiquitin-ligase MID1 and protein phosphatase 2A (PP2A) regulates the nuclear localization and transcriptional activity of GLI3, a transcriptional effector molecule of SHH, in cancer cell lines with autonomously activated SHH signalling. However, the exact molecular mechanisms that mediate the interaction between MID1 and GLI3 remained unknown. Here, we show that MID1 catalyses the ubiquitination and proteasomal cleavage of the GLI3-regulator Fu. Our data…
Isolation and characterization of nuclear microsatellite loci in the northern shrimp, Pandalus borealis
2012
Published version of an article published in the journal: Conservation Genetics Resources. Also available from the publisher at: http://dx.doi.org/10.1007/s12686-011-9486-4 We developed and characterized 20 microsatellite primer loci for the northern shrimp Pandalus borealis. All 20 loci were polymorphic with number of alleles ranging from 3 to 36 and with observed heterozygosity between 0.04 and 0.93. In addition, we tested the utility of these markers in three related shrimp species, P. montagui, Atlantopandalus propinqvus and Dichelopandalus bonnieri. These new markers will prove useful in the identification of stock structure and hence, assessment of the commercially important species P…
Mouse CSB protein is important for gene expression in the presence of a single-strand break in the non-transcribed DNA strand.
2010
CSB protein is required for strand-specific repair of bulky DNA lesions in transcribed genes and mediates transcription recovery after exposure to DNA-damaging agents. We enzymatically generated DNA single-strand breaks (SSBs) with 3'-OH and 5'-phosphate termini in defined positions of a plasmid-borne gene and measured their effect on transcription in cell lines with different statuses of the Csb gene. A single SSB in the transcribed region of the gene caused significant decrease of gene expression. In all tested cell lines of mouse and human origin, a SSB in the transcribed DNA strand was less harmful for gene expression than a SSB situated in the opposing DNA strand. CSB deficiency exhibi…
H2-M polymorphism in mice susceptible to collagen-induced arthritis involves the peptide binding groove.
1996
The ability to develop type II collagen (CII)-induced arthritis (CIA) in mice is associated with the major histocompatibilityI-A gene and with as yet poorly defined regulatory molecules of the major histocompatibility complex (MHC) class II antigen processing and presentation pathway. H2-M molecules are thought to be involved in the loading of antigenic peptides into the MHC class II binding cleft. We sequencedH2-Ma, H2-Mb1, andH2-Mb2 genes from CIA-susceptible and-resistant mouse strains and identified four differentMa andMb2 alleles and three differentMb1 alleles defined by polymorphic residues within the predicted peptide binding groove. Most CIA-resistant mouse strains share commonMa, M…
A global DNA repair mechanism involving the Cockayne syndrome B (CSB) gene product can prevent the in vivo accumulation of endogenous oxidative DNA b…
2002
The Cockayne syndrome B (CSB) gene product is involved in the repair of various types of base modifications in actively transcribed DNA sequences. To investigate its significance for the repair of endogenous oxidative DNA damage, homozygous csb(-/-)/ogg1(-/-) double knockout mice were generated. These combine the deficiency of CSB with that of OGG1, a gene coding for the mammalian repair glycosylase that initiates the base excision repair of 7,8-dihydro-8-oxoguanine (8-oxoG). Compared to ogg1(-/-) mice, csb(-/-)/ogg1(-/-) mice were found to accumulate with age severalfold higher levels of oxidited purine modifications in hepatocytes, splenocytes and kidney cells. In contrast, the basal (ste…
Molecular Effects of the CTG Repeats in Mutant Dystrophia Myotonica Protein Kinase Gene
2008
Myotonic Dystrophy type 1 (DM1) is a multi-system disorder characterized by muscle wasting, myotonia, cardiac conduction defects, cataracts, and neuropsychological dysfunction. DM1 is caused by expansion of a CTG repeat in the 3 untranslated region (UTR) of the Dystrophia Myotonica Protein Kinase (DMPK) gene. A body of work demonstrates that DMPK mRNAs containing abnormally expanded CUG repeats are toxic to several cell types. A core mechanism underlying symptoms of DM1 is that mutant DMPK RNA interferes with the developmentally regulated alternative splicing of defined pre-mRNAs. Expanded CUG repeats fold into ds(CUG) hairpins that sequester nuclear proteins including human Muscleblind-lik…
Psicothema
2009
Resumen tomado de la publicación El objetivo de este estudio es comprobar la efectividad de un programa basado en una metodología lúdico-narrativa para consolidar los contenidos y competencias matemáticas en Educación Infantil en comparación con otras metodologías utilizadas actualmente. Para ello, se seleccionaron 100 niños de 5-6 años que fueron divididos en tres condiciones experimentales: la metodología tradicional basada en fichas de trabajo (centros de interés); una metodología que incluye elementos visuoperceptivos de estimulación temprana y manipulativos, y una metodología que incluye componentes lúdicos y narrativos. Los instrumentos de evaluación utilizados han sido una prueba cri…
Metonymy in Spanish and American parliamentary speeches: Obama’s State of the Union Address versus Rajoy’s State of the Nation Address
2018
This study attempts to analyze the metonymies used in the Economy section of two equivalent parliamentary speeches: the 2015 State of the Union Address in the US and the 2015 State of the Nation Debate in Spain, which belong to two different debate traditions. The present study aims at answering the following research questions:What metonymies do President Obama and Prime Minister Rajoy use in their American and Spanish parliamentary speeches in an attempt to convince the public of economic victory?What are the similarities and differences between the role of metonymy in shaping public opinion about economic recovery in America and Spain in both speeches?To answer these questions, we use th…
Riflessioni sulla struttura del contratto autonomo e della polizza fideiussoria
2012
Il saggio si propone di affermare una linea di metodo per la quale l'interprete che si confronti con le molteplici e multiformi sfaccettature che la prassi mercantile, per soddisfare i bisogni del commercio, imprime alle operazioni contrattuali, deve sempre costantemente rivedere le precedenti acquisizioni, per saggiarne la tenuta di fronte all'evoluzione dei traffici. Allo scopo si sofferma su due ipotesi di scuola ripetutamente affrontate dalla dottrina, talora con esiti davvero pregevoli: il contratto autonomo di garanzia e le polizze fideiussorie.