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Cloning of the cDNA encoding mast cell tryptase of Mongolian gerbil, Meriones unguiculatus, and its preferential expression in the intestinal mucosa

机译:蒙古沙鼠Meriones unguiculatus肥大细胞类胰蛋白酶的cDNA的克隆及其在肠粘膜中的优先表达

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pBy using the combination of reverse-transcription PCR and rapid amplification of cDNA ends methods, a cDNA encoding mast cell tryptase was successfully cloned from the small intestine of Mongolian gerbil, Meriones unguiculatus, infected with Nippostrongylus brasiliensis. The cDNA was 1219 bp long including 810 bp of an open reading frame. Based on the deduced amino acid sequences of known mast cell tryptases of other species, the gerbil mast cell tryptase (gMCT) was highly similar to mouse mast cell protease (mMCP)-7, and seems to be translated as a prepro-enzyme with 25 amino acids of signal and activation peptides and 245 amino acids of mature enzyme. The gMCT mRNA was preferentially transcribed in the intestinal mucosa and to a far lesser extent in the connective tissue such as skin and tongue. Moreover, kinetic study after infection revealed that the amount of gMCT mRNA in the small intestine correlated well with the degree of intestinal mastocytosis. Throughout the course of infection, enzyme-histochemically detectable tryptase activity was limited to mucosal mast cells. Since mucosal mast cells of other rodents, including mice and rats, do not express tryptases, this is the first report of rodent mast cell tryptase expressed in the intestinal mucosa./p
机译:>通过逆转录PCR和cDNA末端快速扩增相结合的方法,成功地从感染了巴西夜蛾的蒙古沙鼠小肠Meriones unguiculatus的小肠中成功克隆了编码肥大细胞类胰蛋白酶的cDNA。 cDNA长1219 bp,其中包括810 bp的开放阅读框。根据推导的其他物种肥大细胞类胰蛋白酶的氨基酸序列,沙鼠肥大细胞类胰蛋白酶(gMCT)与小鼠肥大细胞类蛋白酶(mMCP)-7非常相似,并且似乎可以翻译成25种前原酶信号和激活肽的氨基酸和成熟酶的245个氨基酸。 gMCT mRNA在肠粘膜中优先转录,在结缔组织(如皮肤和舌头)中转录的程度要小得多。此外,感染后的动力学研究表明,小肠中gMCT mRNA的量与肠肥大程度密切相关。在整个感染过程中,酶组织化学可检测的类胰蛋白酶活性仅限于粘膜肥大细胞。由于其他啮齿动物(包括小鼠和大鼠)的粘膜肥大细胞不表达类胰蛋白酶,因此这是首次在肠粘膜中表达啮齿动物肥大细胞类胰蛋白酶的报道。

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