合成生物学视角下的基因功能探索与酵母工程菌株文库构建

章益蜻, 刘高雯

Exploration of gene functions and library construction for engineering strains from a synthetic biology perspective

ZHANG Yiqing, LIU Gaowen

图3 条件等位基因文库（本图由biorender绘制） ［（a）温度敏感型（temperature sensitive，TS）等位基因中，目的基因ORF（橘色片段）被替换成相应的温敏突变基因TS mutant（深橘色片段）并携带一个抗性筛选标签KanMX（黄色片段）。Chr.DNA：染色体DNA。（b）启动子替换策略中，携带抗性筛选标签KanMX（黄色片段）的诱导性启动子TetO（绿色片段）被插入目的基因ORF（橘色片段）的起始密码子上游。Chr.DNA：染色体DNA。（c）通过mRNA扰动降低蛋白丰度的DAmP策略中，目的基因ORF（橘色片段）的3′UTR区被插入一个抗性标签KanR（黄色片段）。Chr.DNA：染色体DNA］

Fig. 3 Construction of conditional allele libraries (created by biorender) [(a) In the temperature-sensitive (TS) allele strategy, the ORF (orange fragment) of target gene is replaced with a corresponding TS mutant (dark orange fragment) and tagged with a KanMX resistance marker (yellow fragment). Chr.DNA: chromosomal DNA.(b) In the promoter replacement strategy, an inducible promoter (green fragment) and a KanMX resistance marker (yellow fragment) is inserted upstream of the start codon of the target gene ORF (orange fragment). Chr.DNA: chromosomal DNA.(c) In the DAmP (Decreased Abundance by mRNA Perturbation) strategy, the 3′UTR of the target gene ORF (orange fragment) is disrupted by insertion of a KanR resistance cassette (yellow fragment) to reduce transcript stability and protein abundance. Chr.DNA: chromosomal DNA.]