title: mazF-mediated deletion system for large-scale genome engineering in Saccharomyces cerevisiae.
abstract: A new mazF-based strategy for large-scale and scarless genome rearrangements in Saccharomyces cerevisiae was developed. We applied this method to delete designed internal (26.5 kbp) and terminal (28.9 kbp) regions located on the left arm of the chromosome XI of S. cerevisiae BY4741. The number of transformants was increased by one order of magnitude and about 90% of tested colonies were desired integrants using in vivo assembled deletion cassette containing longer flanking homology. Compared to conventional URA3 marker, in the counter-selection process, the new system generated 2-13 folds more colonies and the ratio of deletant was simultaneously elevated by 20-24%.
authors.affiliation: Department of Microbiology, College of Life Sciences, Nankai University, Tianjin, China. Electronic address: nk3mlql@gmail.com.
authors.forename: Quanli
authors.initials: Q
authors.lastname: Liu
authors.affiliation: Department of Microbiology, College of Life Sciences, Nankai University, Tianjin, China. Electronic address: kalywyz@sina.com.
authors.forename: Yuzhen
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Hongxun Biotechnologies Ltd, Biobay A2-224, Suzhou, China. Electronic address: ping.yang@incelligen.com.cn.
authors.forename: Ping
authors.initials: P
authors.lastname: Yang
authors.affiliation: Department of Microbiology, College of Life Sciences, Nankai University, Tianjin, China. Electronic address: mingxiuzhang@sina.com.
authors.forename: Xiuming
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Microbiology, College of Life Sciences, Nankai University, Tianjin, China. Electronic address: baiyl@nankai.edu.cn.
authors.forename: Yanling
authors.initials: Y
authors.lastname: Bai
authors.affiliation: Department of Microbiology, College of Life Sciences, Nankai University, Tianjin, China. Electronic address: xhj1971@gmail.com.
authors.forename: Haijin
authors.initials: H
authors.lastname: Xu
authors.affiliation: Department of Microbiology, College of Life Sciences, Nankai University, Tianjin, China; The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Nankai University, Tianjin, China. Electronic address: qiaomq@nankai.edu.cn.
authors.forename: Mingqiang
authors.initials: M
authors.lastname: Qiao
mesh_terms: D004722:Endoribonucleases / Q000378:metabolism*; D017353:Gene Deletion; D005818:Genetic Engineering; D005821:Genetic Techniques; D016681:Genome, Fungal; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D017384:Sequence Deletion
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D004722:Endoribonucleases
keywords: Counter-selection; Large-scale deletion; Saccharomyces cerevisiae; URA3/5-FOA; mazF