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携带有核酸水解 3D8 scFv 的大肠杆菌在 RNA 病毒感染期间的存活情况。

Survival of Escherichia coli harboring nucleic acid-hydrolyzing 3D8 scFv during RNA virus infection.

机构信息

Bio-Evaluation Center, Korea Research Institute of Bioscience and Biotechnology, Cheongju, Chungcheongbuk-do 28116, South Korea.

Bio-Evaluation Center, Korea Research Institute of Bioscience and Biotechnology, Cheongju, Chungcheongbuk-do 28116, South Korea; Department of Food Science and Technology, College of Agriculture and Life Sciences, Chungnam National University, Daejeon 34134, South Korea.

出版信息

Regul Toxicol Pharmacol. 2018 Apr;94:286-292. doi: 10.1016/j.yrtph.2018.02.012. Epub 2018 Feb 24.

Abstract

Previously, Escherichia coli harboring the codon-optimized 3D8scFv gene (E. coli 3D8scFv) was developed as a feed additive for use in preventing norovirus infection. Here, we evaluated whether the 3D8scFv gene affects the colonization of E coli when E. coli 3D8scFv passes through the mouse gastrointestinal tract. To determine the colonization ability of E. coli 3D8scFv, E. coli cells with or without the 3D8scFv gene were fed to mice. Total DNA was extracted from the animals' stools, stomach, small intestine and colon. All samples were amplified using 3D8scFv gene-specific primer sets. E. coli 3D8scFv begins to be excreted 1 h after feeding and that all E. coli 3D8scFv cells were excreted between 12 and 24 h after the last feeding of the cells. The previously measured gastrointestinal transit time of the mice was between 8 h and 22 h. The results of this study therefore show that E. coli 3D8scFv cannot colonize the gastrointestinal tracts of mice. In addition, if the purified 3D8 scFv protein is used as a feed additive, any associated E. coli 3D8scFv bacteria will not colonize the gastrointestinal tracts of the livestock. Thus, this feed additive meets the safety assessment criteria for the commercial use of bacteria.

摘要

先前,我们构建了携带密码子优化的 3D8scFv 基因的大肠杆菌(E. coli 3D8scFv)作为饲料添加剂,用于预防诺如病毒感染。在这里,我们评估了 3D8scFv 基因是否会影响通过小鼠胃肠道的大肠杆菌定植。为了确定 E. coli 3D8scFv 的定植能力,我们用携带或不携带 3D8scFv 基因的大肠杆菌细胞喂食小鼠。从动物粪便、胃、小肠和结肠中提取总 DNA。所有样品均使用 3D8scFv 基因特异性引物进行扩增。喂食后 1 小时开始排泄 3D8scFv 基因,最后一次喂食细胞后 12-24 小时内所有 3D8scFv 基因大肠杆菌细胞都被排泄出来。先前测量的小鼠胃肠道转运时间在 8-22 小时之间。因此,本研究结果表明,E. coli 3D8scFv 不能定植于小鼠的胃肠道。此外,如果将纯化的 3D8 scFv 蛋白用作饲料添加剂,任何相关的 E. coli 3D8scFv 细菌都不会定植于家畜的胃肠道。因此,这种饲料添加剂符合细菌商业用途的安全评估标准。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0584/7115797/150f1e19cc7f/gr1_lrg.jpg

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