Multigene Pathway Engineering with Regulatory Linkers (M-PERL)

详细信息    查看全文

• 作者：Duo Liu ; Hong Liu ; Bing-Zhi Li ; Hao Qi ; Bin Jia ; Xiao Zhou ; Hao-Xing Du ; Wei Zhang ; Ying-Jin Yuan
• 刊名：ACS Synthetic Biology
• 出版年：2016
• 出版时间：December 16, 2016
• 年：2016
• 卷：5
• 期：12
• 页码：1535-1545
• 全文大小：730K
• ISSN：2161-5063

文摘

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.