دورية أكاديمية

A toolbox for genetic manipulation in intestinal Clostridium symbiosum

التفاصيل البيبلوغرافية
العنوان: A toolbox for genetic manipulation in intestinal Clostridium symbiosum
المؤلفون: Pengjie Yang, Jinzhong Tian, Lu Zhang, Hui Zhang, Gaohua Yang, Yimeng Ren, Jingyuan Fang, Yang Gu, Weihong Jiang
المصدر: Synthetic and Systems Biotechnology, Vol 9, Iss 1, Pp 43-54 (2024)
بيانات النشر: KeAi Communications Co., Ltd., 2024.
سنة النشر: 2024
المجموعة: LCC:Biotechnology
LCC:Biology (General)
مصطلحات موضوعية: Gut Clostridium symbiosum, Toolbox, Gene overexpression, Gene deletion, Gene regulation, Biotechnology, TP248.13-248.65, Biology (General), QH301-705.5
الوصف: Gut microbes are closely related with human health, but remain much to learn. Clostridium symbiosum is a conditionally pathogenic human gut bacterium and regarded as a potential biomarker for early diagnosis of intestinal tumors. However, the absence of an efficient toolbox that allows diverse genetic manipulations of this bacterium limits its in-depth studies. Here, we obtained the complete genome sequence of C. symbiosum ATCC 14940, a representative strain of C. symbiosum. On this basis, we further developed a series of genetic manipulation methods for this bacterium. Firstly, following the identification of a functional replicon pBP1 in C. symbiosum ATCC 14940, a highly efficient conjugative DNA transfer method was established, enabling the rapid introduction of exogenous plasmids into cells. Next, we constructed a dual-plasmid CRISPR/Cas12a system for genome editing in this bacterium, reaching over 60 % repression for most of the chosen genes as well as efficient deletion (>90 %) of three target genes. Finally, this toolbox was used for the identification of crucial functional genes, involving growth, synthesis of important metabolites, and virulence of C. symbiosum ATCC 14940. Our work has effectively established and optimized genome editing methods in intestinal C. symbiosum, thereby providing strong support for further basic and application research in this bacterium.
نوع الوثيقة: article
وصف الملف: electronic resource
اللغة: English
تدمد: 2405-805X
Relation: http://www.sciencedirect.com/science/article/pii/S2405805X23001126; https://doaj.org/toc/2405-805X
DOI: 10.1016/j.synbio.2023.12.005
URL الوصول: https://doaj.org/article/f2acde9e8ed441268fb2b545fbfc58ce
رقم الأكسشن: edsdoj.f2acde9e8ed441268fb2b545fbfc58ce
قاعدة البيانات: Directory of Open Access Journals
الوصف
تدمد:2405805X
DOI:10.1016/j.synbio.2023.12.005