文献类型：期刊文献

英文题名：ETfinder: harnessing conserved C-terminal tails of single-stranded DNA-binding proteins for mining and engineering RecET systems in non-model microbial chassis

作者：Lv, Dongyuan[1,2];Liang, Mindong[1,2];Gu, Yiyang[1,2];Zhu, Xiangying[3];Wang, Hailong[4];Xia, Menglei[5];Hao, Jinkang[5];Li, Youyuan[1,2];Wang, Weishan[6];Bai, Linquan[7,8];Cheng, Jiagao[1,2,3];Zhang, Lixin[1,2];Tan, Gao-Yi[1,2]

机构：[1]East China Univ Sci & Technol ECUST, State Key Lab Bioreactor Engn SKLBE, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol ECUST, Sch Biotechnol, Shanghai 200237, Peoples R China;[3]East China Univ Sci & Technol, Sch Pharm, Shanghai Key Lab Chem Biol, Shanghai 200237, Peoples R China;[4]Shandong Univ, Sch Life Sci, State Key Lab Microbial Technol, Jinan 250100, Peoples R China;[5]Tianjin Univ Sci & Technol, Coll Biotechnol, Metab & Fermentat Proc Control, Minist Educ, Tianjin 300457, Peoples R China;[6]Chinese Acad Sci, State Key Lab Microbial Resources, Inst Microbiol, Beijing 100101, Peoples R China;[7]Shanghai Jiao Tong Univ, State Key Lab Microbial Metab, Shanghai 200240, Peoples R China;[8]Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, Shanghai 200240, Peoples R China

年份：2026

卷号：54

期号：15

外文期刊名：NUCLEIC ACIDS RESEARCH

收录：;Scopus(收录号：2-s2.0-105047705044);WOS:【SCI-EXPANDED(收录号:WOS:001853386400001)】；

基金：This work was financially supported by grants from the National Key Research and Development Program of China (grant no. 2020YFA0907304 to T.G.-Y.); the National Natural Science Foundation of China (grant nos. 32121005 to Z.L., and 32370064 and 31870040 to T.G.-Y.); the Shanghai Oriental Talents Plan (BJZH2024085 to T.G.-Y); the Fundamental Research Funds for the Central Universities (to T.G.-Y.) and the 2023 Double World-Class Project - Key Program for Intelligent Biomanufacturing (to Z.L.). Funding to pay the Open Access publication charges for this article was provided by the National Key Research and Development Program of China (2020YFA0907304).

语种：英文

摘要：Nonmodel microorganisms offer substantial potential as next-generation microbial chassis (NGMCs), yet most lack efficient and broadly transferable genome-editing systems. Here we present ETfinder, a framework that uses the conserved C-terminal tail of host single-stranded DNA-binding proteins (SSB-Ct) as a biochemical constraint to guide the discovery of RecET recombineering systems. Applied to Rhodobacter sphaeroides, ETfinder identified 91 candidates from 18 841 alpha-proteobacterial genomes, and all five experimentally tested RecT homologs supported measurable double-stranded DNA (dsDNA) recombineering, with the Paracoccaceae SJ630 system reaching 8.9 & times; 10 & sup2; colony-forming units (CFU) per mu g of dsDNA and 100% editing accuracy. Testing in Halomonas further showed that RecT proteins from evolutionarily distant taxa remain functional within the same halophilic chassis, indicating that SSB-Ct-guided selection enriches for portable recombination modules beyond phylogenetic proximity. To facilitate broad adoption, we compiled 25 529 RecT-SSB pairs into a curated database and implemented ETfinder as a standalone, locally deployable toolkit for mining, ranking, and phylogenetic visualization. This framework prioritizes high-compatibility homologs, reduces experimental screening burden, and expands the accessible genome-editing toolbox for NGMCs. ETfinder is freely available at https://github.com/lvdongyuan/ETfinder.