PrimePCR™ Probe Assay: Plec, Mouse

RT

Real-time PCR probe assay designed for gene expression analysis. Probe assays consist of unlabeled PCR primers and a dual labeled fluorescent probe.

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Assay Information

Technology:   qPCR
Assay Type:   Probe
Application:   Gene Expression
Unique Assay ID:   qMmuCIP0030043
Assay Design:   Intron-spanning
Chromosome Location:   15:76186740-76187754question
Amplicon Length:   112
Splice Variants Targeted:   ENSMUST00000074834 ENSMUST00000023226 ENSMUST00000072692 ENSMUST00000169438 ENSMUST00000169714 ENSMUST00000169108 ENSMUST00000076442 ENSMUST00000171562 ENSMUST00000165210 ENSMUST00000171634 ENSMUST00000089610 ENSMUST00000071869 ENSMUST00000073418 ENSMUST00000054449 ENSMUST00000080857

Gene Information

Plectin is a prominent member of an important family of structurally and in part functionally related proteins termed plakins or cytolinkers that are capable of interlinking different elements of the cytoskeleton. Plakins with their multi-domain structure and enormous size not only play crucial roles in maintaining cell and tissue integrity and orchestrating dynamic changes in cytoarchitecture and cell shape but also serve as scaffolding platforms for the assembly positioning and regulation of signaling complexes (reviewed in PMID: 9701547 11854008 17499243). Plectin is expressed as several protein isoforms in a wide range of cell types and tissues from a single gene located on chromosome 8 in humans (PMID: 8633055 8698233). Until 2010 this locus was named plectin 1 (symbol PLEC1 in human; Plec1 in mouse and rat) and the gene product had been referred to as "hemidesmosomal protein 1" or "plectin 1 intermediate filament binding 500kDa". These names were superseded by plectin. The plectin gene locus in mouse on chromosome 15 has been analyzed in detail (PMID: 10556294 14559777) revealing a genomic exon-intron organization with well over 40 exons spanning over 62 kb and an unusual 5' transcript complexity of plectin isoforms. Eleven exons (1-1j) have been identified that alternatively splice directly into a common exon 2 which is the first exon to encode plectin's highly conserved actin binding domain (ABD). Three additional exons (-1 0a and 0) splice into an alternative first coding exon (1c) and two additional exons (2alpha and 3alpha) are optionally spliced within the exons encoding the acting binding domain (exons 2-8). Analysis of the human locus has identified eight of the eleven alternative 5' exons found in mouse and rat (PMID: 14672974); exons 1i 1j and 1h have not been confirmed in human. Furthermore isoforms lacking the central rod domain encoded by exon 31 have been detected in mouse (PMID:10556294) rat (PMID: 9177781) and human (PMID: 11441066 10780662 20052759). It has been shown that the short alternative amino-terminal sequences encoded by the different first exons direct the targeting of the various isoforms to distinct subcellular locations (PMID: 14559777). As the expression of specific plectin isoforms was found to be dependent on cell type (tissue) and stage of development (PMID: 10556294 12542521 17389230) it appears that each cell type (tissue) contains a unique set (proportion and composition) of plectin isoforms as if custom-made for specific requirements of the particular cells. Concordantly individual isoforms were found to carry out distinct and specific functions (PMID: 14559777 12542521 18541706). In 1996 a number of groups reported that patients suffering from epidermolysis bullosa simplex with muscular dystrophy (EBS-MD) lacked plectin expression in skin and muscle tissues due to defects in the plectin gene (PMID: 8698233 8941634 8636409 8894687 8696340). Two other subtypes of plectin-related EBS have been described: EBS-pyloric atresia (PA) and EBS-Ogna. For reviews of plectin-related diseases see PMID: 15810881 19945614. Mutations in the plectin gene related to human diseases should be named based on the position in NM_000445 (human variant 1 isoform 1c) unless the mutation is located within one of the other alternative first exons in which case the position in the respective Reference Sequence should be used. [provided by RefSeq Aug 2011]

Gene Symbol:   Plec
Gene Name:   Plectin
Aliases:   AA591047, AU042537, EBS1, PCN, PLTN, Plec1
RefSeq:   NC_000081.6 NT_039621.8
Ensembl:   ENSMUSG00000022565
Entrez:   18810
Chromosome Mapping:   15 D3|15 44.0 cM

The below validation information is for the Primer Pair only   Download Validation Data (.pdf)


Products used to generate validation data:

Real-Time PCR Instrument CFX384 Real-Time PCR Detection System
Reverse Transcription Reagent iScript™ Advanced cDNA Synthesis Kit for RT-qPCR
Real-Time PCR Supermix SsoAdvanced™ SYBR® Green Supermix
Experimental Sample qPCR Mouse Reference Total RNA

Summary Data:

R2 0.999700
y-intercept 35.210000
Efficiency 97

Amplification Plot
Amplification of cDNA generated from universal RNA.

Amplification of cDNA generated from universal RNA.

Melt Peak
Melt curve analysis of above amplification.

Melt curve analysis of above amplification.

Standard Curve
Standard curve generated using 20 million copies of template diluted 10 fold to 20 copies.

Standard curve generated using 20 million copies of template diluted 10 fold to 20 copies.

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