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首页> 外文期刊>Biological chemistry >Studies of intestinal morphology and cathepsin B expression in a transgenic mouse aiming at intestine-specific expression of Cath B-EGFP.
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Studies of intestinal morphology and cathepsin B expression in a transgenic mouse aiming at intestine-specific expression of Cath B-EGFP.

机译:旨在针对Cath B-EGFP的肠特异性表达的转基因小鼠中肠形态和组织蛋白酶B表达的研究。

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摘要

Cathepsin B has been shown to not only reside within endo-lysosomes of intestinal epithelial cells, but it was also secreted into the extracellular space of intestinal mucosa in physiological and pathological conditions. In an effort to further investigate the function of this protease in the intestine, we generated a transgenic mouse model that would enable us to visualize the localization of cathepsin B in vivo. Previously we showed that the A33-antigen promoter could be successfully used in vitro in order to express cathepsin B-green fluorescent protein chimeras in cells that co-expressed the intestine-specific transcription factor Cdx1. In this study an analog approach was used to express chimeric cathepsin B specifically in the intestine of transgenic animals. No overt phenotype was observed for the transgenic mice that reproduced normally. Biochemical and morphological studies confirmed that the overall intestinal phenotype including the structure and polarity of this tissue as well as cell numbers and differentiation states were not altered in the A33-CathB-EGFP mice when compared to wild type animals. However, transgenic expression of chimeric cathepsin B could not be visualized because it was not translated in situ although the transgene was maintained over several generations.
机译:已经证明组织蛋白酶B不仅存在于肠上皮细胞的内溶酶体内,而且在生理和病理条件下也被分泌到肠粘膜的细胞外空间。为了进一步研究这种蛋白酶在肠中的功能,我们生成了一个转基因小鼠模型,该模型可使我们可视化组织蛋白酶B在体内的定位。以前,我们表明A33抗原启动子可以成功地在体外用于在共同表达肠特异性转录因子Cdx1的细胞中表达组织蛋白酶B-绿色荧光蛋白嵌合体。在这项研究中,使用一种模拟方法在转基因动物的肠道中特异性表达嵌合组织蛋白酶B。对于正常繁殖的转基因小鼠没有观察到明显的表型。生化和形态学研究证实,与野生型动物相比,A33-CathB-EGFP小鼠的肠道整体表型(包括该组织的结构和极性以及细胞数量和分化状态)没有改变。然而,嵌合组织蛋白酶B的转基因表达无法观察到,因为尽管转基因已经维持了好几代,但它并未被原位翻译。

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