Biomedical subjects

N Peyrol

Publications and source records attributed to N Peyrol.

3 recordsLinked to original sources

PubMed1997

Cloning and initial characterization of human and mouse Spot 14 genes.

The intricate regulation of Spot 14 expression in rat lipogenic tissues has provided a useful tool in studying nutritional and hormonal factors involved in transcription. To gain insight into its function and its possible involvement in human lipid disorders, we cloned human and mouse Spot 14 genes that shared with the rat gene a strong homology concerning the deduced amino acid sequence (81 and 94%, respectively) as well as the promoter region. The mouse promoter was characterized by transfection studies, while quantitative RT-PCR and in situ hybridization experiments showed that Spot 14 is expressed in human liver and, at a high level, in multiple symmetric lipomatosis nodules.

Amino Acid Sequence↗

PubMed1996

Quantitative multistandard RT-PCR assay using interspecies polymorphism.

The use of RT-PCR to quantify mRNA is often compromised by the variability of reverse-transcription and amplification reactions as well as by the difficulty of assessing the amount and/or the integrity of input RNA. Use of a competitor RNA or the coamplification of an endogenous standard are widespread methods of monitoring these steps. Taking advantage of both sequence conservation between homologous genes in related animal species and interspecific polymorphism, a protocol that may be regarded as a compromise between these two methods is described here. Total RNA samples, extracted from even minute amounts of tissue belonging to a first animal species, were supplemented with a constant amount of total RNA prepared from a second animal species, which thus acts as a multistandard source. The mixture was reverse-transcribed using hexa-random primers. Separate PCRs were then undertaken so that, for each mRNA of interest, products from both origins could be distinguished. Since the ratio between amplified mRNAs is constant in the standard preparation, an accurate normalization in the assay samples of most variations inherent to PCR is obtained. This protocol allows quantification of several mRNAs species, whose amounts may be very different, in a single cDNA preparation.

Actins↗

PubMed1991

Production of rat Spot14 protein in Escherichia coli.

Hormonal, nutritional and developmental factors modulate, in rat lipogenic tissues, the transcription of the mRNA coding for a protein of unknown function, called Spot14 (S14). The corresponding protein has never been purified from tissues. In this paper, we describe the production of S14 in Escherichia coli. In the absence of available antibodies (Ab) directed against S14 protein, our strategy was to produce this protein by constructing two different recombinant expression vectors. The first recombinant plasmid produced a S14::protein A fusion which was easily purified and then rabbit Ab could be raised against it. The second expression vector directly produced S14. This expression was demonstrated by specific binding of polyclonal Ab directed against the fusion protein. These Ab also recognized a rat-liver protein sharing characteristics of S14.

Animals↗