Search results for "primer"
showing 10 items of 530 documents
A Mutation With Major Effects on Drosophila melanogaster Sex Pheromones
2005
0016-6731 (Print) Comparative Study Journal Article Research Support, Non-U.S. Gov't; Sex pheromones are intraspecific chemical signals that are crucial for mate attraction and discrimination. In Drosophila melanogaster, the predominant hydrocarbons on the cuticle of mature female and male flies are radically different and tend to stimulate or inhibit male courtship, respectively. This sexual difference depends largely upon the number of double bonds (one in males and two in females) added by desaturase enzymes. A mutation was caused by a PGal4 transposon inserted in the desat1 gene that codes for the desaturase crucial for setting these double bonds. Homozygous mutant flies produced 70-90%…
Altered splicing pattern of TACC1 mRNA in gastric cancer
2002
Abstract Transforming acidic coiled-coil ( TACC ) proteins are centrosome and microtubule-associated proteins that are essential for mitotic spindle function. We identified TACC1 as an immunogenic protein and a potential tumor antigen by applying serological identification of antigens by recombinant expression cloning (SEREX) technique to screen a gastric cancer cDNA library. The 5′RLM-RACE and reverse transcriptase polymerase chain reaction analyses revealed at least six different transcript variants of TACC1 with variable transcription start sites and alternative exon usage (designated TACC1-A–TACC1-F ). All transcripts differ in their 5′ ends but share an identical 3′ region encoding coi…
The voice of a heterodox authority on love in the First Francoism. Amparo Rivelles’ sentimental advice columns
2021
Entre 1948 y 1952, el semanario cinematográfico Primer plano publicó un consultorio sentimental firmado por la actriz Amparo Rivelles. La estrella española más popular del momento se convertía así en consejera sobre la moral, el noviazgo y los usos amorosos entre los jóvenes; una cuestión que el Régimen consideraba como una pieza fundamental en la regulación de las relaciones de género. Se trata de un consultorio singular, alejado de la gravedad, el dramatismo y la grandilocuencia de otros, surgidos en diversos medios periodísticos a raíz de la buena acogida de público que estas secciones gozaron durante aquellos años. El presente artículo analiza las implicaciones de unos textos en los que…
Rol de las familias. Escuelas y docentes en las interacciones entre tecnología digital y primera Infancia y sus posibles efectos
2022
En este capítulo se pretende describir los usos e influencia de la tecnología centrados en la Primera Infancia, tanto en la escuela como en el hogar. Se reflexiona sobre la interacción entre infancia, tecnología, familias y escuela, partiendo de la Teoría ecológica de sistemas y en el marco de la sociedad digital del siglo XXI. This chapter aims to describe the uses and influence of technology focused on Early Childhood, both at school and at home. It reflects on the interaction between childhood, technology, families and school, based on the ecological theory of systems and within the framework of the digital society of the 21st century. Se destaca el papel relevante que tiene la comunicac…
Rapid PCR-based test for identifying Candida albicans by using primers derived from the pH-regulated KER1 gene.
2006
A PCR-based method in combination with a simple, reliable and inexpensive DNA extraction procedure for rapid detection of Candida albicans clinical isolates is described here. The extraction protocol is based on a combination of chemical (NaOH and detergents) and physical (boiling) treatments, thus avoiding many of the problems inherent in the currently available DNA extraction protocols (basically the use of expensive and/or toxic chemical reagents), and may be useful for daily clinical routine. The PCR-based system described here uses a single pair of primers (SC1F and SC1R) deduced from the C. albicans-specific KER1 gene sequence. These primers amplify a 670-bp fragment of the KER1 gene.…
Role of Human Sec63 in Modulating the Steady-State Levels of Multi-Spanning Membrane Proteins
2012
The Sec61 translocon of the endoplasmic reticulum (ER) membrane forms an aqueous pore, allowing polypeptides to be transferred across or integrated into membranes. Protein translocation into the ER can occur co- and posttranslationally. In yeast, posttranslational translocation involves the heptameric translocase complex including its Sec62p and Sec63p subunits. The mammalian ER membrane contains orthologs of yeast Sec62p and Sec63p, but their function is poorly understood. Here, we analyzed the effects of excess and deficit Sec63 on various ER cargoes using human cell culture systems. The overexpression of Sec63 reduces the steady-state levels of viral and cellular multi-spanning membrane …
Direct quantification of cell-free, circulating DNA from unpurified plasma.
2014
Cell-free DNA (cfDNA) in body tissues or fluids is extensively investigated in clinical medicine and other research fields. In this article we provide a direct quantitative real-time PCR (qPCR) as a sensitive tool for the measurement of cfDNA from plasma without previous DNA extraction, which is known to be accompanied by a reduction of DNA yield. The primer sets were designed to amplify a 90 and 222 bp multi-locus L1PA2 sequence. In the first module, cfDNA concentrations in unpurified plasma were compared to cfDNA concentrations in the eluate and the flow-through of the QIAamp DNA Blood Mini Kit and in the eluate of a phenol-chloroform isoamyl (PCI) based DNA extraction, to elucidate the D…
Genomic Structure and in Vivo Expression of the Human Organic Anion Transporter 1 (hOAT1) Gene
2000
The human organic anion transporter 1 (hOAT1) plays a key role in the secretion of an array of potentially toxic organic anions including many clinically important drugs. Here we report on the genomic cloning of hOAT1. A human genomic library was used for screening of a PAC (P1 artificial chromosome) clone applying PCR techniques. Sequencing of several restriction subclones and of a PCR-generated clone revealed that the hOAT1 gene spans 8.2 kb and is composed of 10 exons divided by 9 introns. RT-PCR studies in a human kidney specimen led to the detection of two new splice variants, hOAT1-3 and hOAT1-4, showing a 132-bp in-frame deletion. Using fluorescence in situ hybridization (FISH) we ma…
Quantitative RT-PCR measurement of human cytochrome P-450s: application to drug induction studies.
2000
A quantitative RT-PCR assay has been developed that is able to measure the mRNA content of the major human CYPs (1A1, 1A2, 2A6, 2B6, 2C9, 2C19, 2D6, 2E1, 3A4, and 3A5). The technique is highly specific, reproducible, rapid, and sensitive enough to quantitate low and high abundant mRNAs. The PCR primers were selected to specifically match each CYP mRNA, to have a very close annealing temperature, and to render PCR products of similar sizes. The PCR conditions were designed to allow the simultaneous measurement of the various human liver CYPs in a single run. To achieve precise and reproducible quantitation of each cytochrome mRNA, a external standard (luciferase mRNA) is added to the probes …
Different La/SS-B mRNA isoforms are expressed in salivary gland tissue of patients with primary Sjogren's syndrome
1996
Recently we isolated a La/SS-B mRNA isoform from a cDNA library made from peripheral blood lymphocytes of a patient with primary Sjögren's Syndrome. In the La/SS-B mRNA isoform the exon 1 was replaced. The alternative exon was termed exon 1'. Genomic analysis showed that the exon 1' La mRNA was the result of a promoter-switch in combination with alternative splicing. Due to the unusual structure of the exon 1' La/SS-B mRNA, the function and the behaviour under physiological and pathophysiological conditions in tissue of patients with primary Sjögren's syndrome or Systemic Lupus Erythematosus remained obscure. Therefore assays were established allowing a qualitative and quantitative estimati…