合成生物学
生物
DNA
生物制造
计算生物学
模板
质粒
凝聚体
无细胞蛋白质合成
遗传学
基因组
基因
计算机科学
程序设计语言
蛋白质生物合成
作者
Sung Sun Yim,Nathan I Johns,Vincent Noireaux,Harris H. Wang
标识
DOI:10.1021/acssynbio.0c00277
摘要
Recent advances in cell-free systems have opened up new capabilities in synthetic biology from rapid prototyping of genetic circuits and metabolic pathways to portable diagnostics and biomanufacturing. A current bottleneck in cell-free systems, especially those employing non-E. coli bacterial species, is the required use of plasmid DNA, which can be laborious to construct, clone, and verify. Linear DNA templates offer a faster and more direct route for many cell-free applications, but they are often rapidly degraded in cell-free reactions. In this study, we evaluated GamS from λ-phage, DNA fragments containing Chi-sites, and Ku from Mycobacterium tuberculosis for their ability to protect linear DNA templates in diverse bacterial cell-free systems. We show that these nuclease inhibitors exhibit differential protective activities against endogenous exonucleases in five different cell-free lysates, highlighting their utility for diverse bacterial species. We expect these linear DNA protection strategies will accelerate high-throughput approaches in cell-free synthetic biology.
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