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基于重组酶的体内表达技术在猪霍乱沙门氏菌亚种中的应用,用于鉴定小鼠胃肠道中诱导表达的基因。

Application of Recombinase-Based In Vivo Expression Technology to subsp. for Identification of Genes Induced in the Gastrointestinal Tract of Mice.

作者信息

Koguchi Hiroka, Ishigami Natsumi, Sakanaka Mikiyasu, Yoshida Kako, Hiratou Sayaka, Shimada Mina, Fukiya Satoru, Sonoyama Kei, Yokota Atsushi

机构信息

Laboratory of Microbial Physiology, Research Faculty of Agriculture, Hokkaido University, Sapporo 060-8589, Japan.

Laboratory of Food Biochemistry, Research Faculty of Agriculture, Hokkaido University, Sapporo 060-8589, Japan.

出版信息

Microorganisms. 2020 Mar 13;8(3):410. doi: 10.3390/microorganisms8030410.

Abstract

Bifidobacteria are one of the major components in human gut microbiota and well-known as beneficial microbes. However, clarification of commensal mechanisms of bifidobacteria in the intestines is still ongoing, especially in the presence of the gut microbiota. Here, we applied recombinase-based in vivo expression technology (R-IVET) using the bacteriophage P1 Cre/ system to subsp. 105-A ( 105-A) to identify genes that are specifically expressed in the gastrointestinal tract of conventionally raised mice. Oral administration of the genomic DNA library of 105-A to conventionally raised mice resulted in the identification of 73 in vivo-induced genes. Four out of seven tested genes were verified in vivo-specific induction at least in the cecum by quantitative reverse transcription PCR. Although there is still room for improvement of the system, our findings can contribute to expanding our understanding of the commensal behavior of in the gut ecosystem.

摘要

双歧杆菌是人体肠道微生物群的主要组成部分之一,是著名的有益微生物。然而,双歧杆菌在肠道内的共生机制仍在研究中,尤其是在肠道微生物群存在的情况下。在这里,我们应用基于重组酶的体内表达技术(R-IVET),使用噬菌体P1 Cre/系统对105-A亚种(105-A)进行研究,以鉴定在常规饲养小鼠胃肠道中特异性表达的基因。将105-A的基因组DNA文库口服给常规饲养的小鼠,结果鉴定出73个体内诱导基因。通过定量逆转录PCR,在七个测试基因中,有四个至少在盲肠中被证实为体内特异性诱导。尽管该系统仍有改进空间,但我们的研究结果有助于扩大我们对105-A在肠道生态系统中共生行为的理解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/02f3/7143038/a11d12a95e60/microorganisms-08-00410-g001.jpg

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