Cellular reagents for diagnostics and synthetic biology.

Cellular reagents for diagnostics and synthetic biology.
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DOI:
10.1371/journal.pone.0201681
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发表时间:
2018
期刊:
影响因子:
3.7
通讯作者:
Ellington AD
Ellington AD
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Bhadra S;Pothukuchy A;Shroff R;Cole AW;Byrom M;Ellefson JW;Gollihar JD;Ellington AD

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我们已经发现,细菌中酶的过量生产,然后将其冻干,导致可以直接用于进行许多分子生物学反应的“细胞试剂”。我们证明了细胞试剂在各种分子诊断中的使用,例如不降低灵敏度的TaqMan qPCR,以及在合成生物学基础中的使用,例如DNA片段的吉布森组装,其中可以仅基于添加细胞试剂来构建新质粒。细胞试剂大大降低了生产、储存和实施的复杂性和成本,这些特点应有助于在资源贫乏的条件下获得和使用。
We have found that the overproduction of enzymes in bacteria followed by their lyophilization leads to 'cellular reagents' that can be directly used to carry out numerous molecular biology reactions. We demonstrate the use of cellular reagents in a variety of molecular diagnostics, such as TaqMan qPCR with no diminution in sensitivity, and in synthetic biology cornerstones such as the Gibson assembly of DNA fragments, where new plasmids can be constructed solely based on adding cellular reagents. Cellular reagents have significantly reduced complexity and cost of production, storage and implementation, features that should facilitate accessibility and use in resource-poor conditions.
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