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Phylogenetic debugging of a complete human biosynthetic pathway transplanted into yeast

Nucleic Acids Res.. 2020; 
Agmon N1, Temple J1, Tang Z2, Schraink T3, Baron M4, Chen J5, Mita P1, Martin JA1, Tu BP5, Yanai I4, Fenyö D1, Boeke JD1.
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PCR Cloning and Subcloning … NP 001117.2). CDS were designed with appropriate over- hangs and restriction<br> enzyme to allow for cloning us- ing yGG assembly (18). Fragments were synthesized<br> and cloned through Gen9Bio or <b>GenScript</b>. For expression … Get A Quote

摘要

Cross-species pathway transplantation enables insight into a biological process not possible through traditional approaches. We replaced the enzymes catalyzing the entire Saccharomyces cerevisiae adenine de novo biosynthesis pathway with the human pathway. While the 'humanized' yeast grew in the absence of adenine, it did so poorly. Dissection of the phenotype revealed that PPAT, the human ortholog of ADE4, showed only partial function whereas all other genes complemented fully. Suppressor analysis revealed other pathways that play a role in adenine de-novo pathway regulation. Phylogenetic analysis pointed to adaptations of enzyme regulation to endogenous metabolite level 'setpoints' in diverse organisms. Using... More

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