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N Pillet

Publications and source records attributed to N Pillet.

6 recordsLinked to original sources
PubMed1995

Characterization of the promoter region of the mouse Xist gene.

The mouse Xist gene is expressed exclusively from the inactive X chromosome and may be implicated in initiating X inactivation. To better understand the mechanisms underlying the control of Xist expression, we investigated the upstream regulatory region of the mouse Xist promoter. A 1.2-kb upstream region of the Xist gene was sequenced and promoter activity was studied by chloramphenicol acetyltransferase (CAT) assays after transfection in murine XX and XY cell lines. The region analyzed (-1157 to +917 showed no in vitro sex-specific promoter activity. However, a minimal constitutional promoter was assigned to a region from -81 to +1, and a cis element from -41 to -15 regulates promoter activity. We showed that a nuclear factor binds to an element located at -30 to -25 (TTAAAG). A second sequence at -41 to -15 does not act as an enhancer and is unable to confer transcriptional activity to the Xist gene on its own. A third region from -82 to -41 is needed for correct expression. Deletion of the segment -441 to -231 is associated with an increase in CAT activity and may represent a silencer element.

3T3 Cells↗
PubMed1995

An acyclic 5-nitroindazole nucleoside analogue as ambiguous nucleoside.

Acyclic nucleoside analogues with carboxamido- or nitro-substituted heterocyclic bases have been evaluated for their possible use as universal bases in oligodeoxynucleotides. The acyclic moiety endows the constructs with enough flexibility to allow good base stacking. The 5-nitroindazole analogue afforded the most stable duplexes among the acyclic derivatives with the least spread in Tm versus the four natural bases. In spite of the acyclic moiety, stabilities are comparable with those of duplexes incorporating the recently described 5-nitroindole nucleoside analogue, but considerably exceed those for the 3-nitropyrrole analogue.

Base Sequence↗
PubMed1994

[Diagnosis using PCR: the direct approach].

The direct approach in molecular diagnosis proposes evidences of the mutations underlying the investigated diseases. Due to its speed, specificity and low cost, the Polymerase Chain Reaction (PCR) has become the method of choice in most of these analyses. The direct approach can be either positive or negative. In the first case, the diagnosis is based on the presence of a pathognomonic PCR product, while in the latter, it is the absence of such a product that makes the diagnosis possible. It is evident that both methods have to be validated by several control reactions. Examples out of daily practice illustrate various diagnostic areas.

Base Sequence↗
PubMed1994

[Diagnosis using PCR: the indirect approach].

The goal of indirect molecular diagnostic techniques is the detection of a link between a specific allele of a polymorphic genomic marker and a given disease. This technique is used in two specific clinical situations: 1) when the gene is unknown and the search for a mutation or a gene product is impossible 2) when the gene is known but large and several mutations might be present. If none of the known mutations prevails in the local population the systematic and sequential search for every single mutation is not economical. A linkage study is required in those instances. The indirect analysis was up to the nineties based on markers detecting DNA-restriction fragments of various length. By the amplification of microsatellites by PCR the indirect approach has brought enormous progress. Together with the mapping by the Human Genome Project it will progress further. Although indirect molecular methodology can often avoid extended work it is only applicable to families with a member already affected by the disease. DNA from this individual is needed for the detection of a link of one allele to the disease. This means necessarily that the disease must be inherited and that the diagnosis is certain. Presymptomatic molecular diagnosis is illustrated by the analysis of a pedigree with familial adenomatous polyposis by microsatellite PCR techniques.

Adenomatous Polyposis Coli↗