Characterization of tree shrew APP gene and identification of its alternatively spliced variants
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    Abstract:

    Objective The aim of this study was to differentiate and identify all the variants of APP mRNA in tree shrew, describe the characteristics of APP genes, and determine its distribution in different tissues. Method Based on the known human APP sequence and predicted tree shrew APP gene, we designed a pair of common specific primer. We extracted total RNA from different tissues respectively, using RT-PCR to get the targeted cDNA. We differentiated the variants by electrophoresis. Finally we recollected the RT-PCR products for sequencing. Combined with the qPCR results, we confirmed the quantitative distribution of the variants in different tissues. Results Our sequencing results showed that the length of the tree shrew APP spliceosome was 3514 bp with a 109 bp 5'-UTR and a 1092 bp3'-UTP. APP genes in the investigated 9 species were highly homologous and conservative, tree shrews and primates show a very close genetic relationship. The tree shrew APP genes shared four domains by 3D modeling. We also confirmed the distribution pattern of 4 spliceosomes derived from exon jumping. All existed APP variants, including APP770, APP695, APP751, and APP677, were all expressed in the lung, kidney and intestine. The highest expression levels were in the lung, muscle, and testicles. Conclusions This study helps to promote the studies of tree shrews as an Alzheimer's disease model and the mechanism of its pathogenesis and drug development.

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History
  • Received:January 04,2017
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  • Online: April 28,2017
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