title: Multigene Pathway Engineering with Regulatory Linkers (M-PERL).
abstract: Multigene pathway engineering usually needs amounts of part libraries on transcriptional and translational regulation as well as mutant enzymes to achieve the optimal part combinations of the target pathways. We report a new strategy for multigene pathway engineering with regulatory linkers (M-PERL) focusing on tuning the transcriptional start site (TSS) of yeast promoters. The regulatory linkers are composed of two homologous ends of two adjacent gene parts for assembly and a central regulatory region between them. We investigated the effect of the homologous end's length on multigene assembly, analyzed the influences of truncated, replaced, and elongated TSS and the adjacent region on promoters, and introduced 5 to 40 random bases of N (A/T/C/G) in the central regulatory region of the linkers which effectively varied the promoter's strengths. The distinct libraries of five regulatory linkers were used simultaneously to assemble and tune all five genes in the violacein synthesis pathway. The gene expressions affected the product profiles significantly, and the recombinants for enhanced single component synthesis and varied composition synthesis were obtained. This study offers an efficient tool to assemble and regulate multigene pathways.
authors.affiliation: Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University , Tianjin, 300072, P. R. China.|SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin) , Tianjin, 300072, P. R. China.
authors.forename: Duo
authors.initials: D
authors.lastname: Liu
authors.affiliation: Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University , Tianjin, 300072, P. R. China.|SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin) , Tianjin, 300072, P. R. China.
authors.forename: Hong
authors.initials: H
authors.lastname: Liu
authors.affiliation: Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University , Tianjin, 300072, P. R. China.|SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin) , Tianjin, 300072, P. R. China.
authors.forename: Bing-Zhi
authors.initials: BZ
authors.lastname: Li
authors.affiliation: Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University , Tianjin, 300072, P. R. China.|SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin) , Tianjin, 300072, P. R. China.
authors.forename: Hao
authors.initials: H
authors.lastname: Qi
authors.affiliation: Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University , Tianjin, 300072, P. R. China.|SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin) , Tianjin, 300072, P. R. China.
authors.forename: Bin
authors.initials: B
authors.lastname: Jia
authors.affiliation: Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University , Tianjin, 300072, P. R. China.|SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin) , Tianjin, 300072, P. R. China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University , Tianjin, 300072, P. R. China.|SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin) , Tianjin, 300072, P. R. China.
authors.forename: Hao-Xing
authors.initials: HX
authors.lastname: Du
authors.affiliation: Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University , Tianjin, 300072, P. R. China.|SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin) , Tianjin, 300072, P. R. China.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University , Tianjin, 300072, P. R. China.|SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin) , Tianjin, 300072, P. R. China.
authors.forename: Ying-Jin
authors.initials: YJ
authors.lastname: Yuan
mesh_terms: D015870:Gene Expression; D015723:Gene Library; D005818:Genetic Engineering / Q000379:methods*; D007211:Indoles / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012045:Regulatory Sequences, Nucleic Acid; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D024363:Transcription Initiation Site
chemical_list: D007211:Indoles; C063155:violacein
keywords: DNA assembly; multigene pathway engineering; regulatory linker; regulatory region; yeast promoter