A facile method for high-throughput co-expression of protein pairs
A facile method for high-throughput co-expression of protein pairs
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DOI：
10.1074/mcp.t400008-mcp200
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发表时间：
2004-09-01
影响因子：
7
通讯作者：
Grayhack, EJ
中科院分区：
文献类型：
--
作者：
Alexandrov, A;Vignali, M;Grayhack, EJ
我们开发了一种方法，通过将61个核苷酸的序列（LINK）整合到质粒中以允许产生串联质粒，从表达不同ORF的其他相同的大肠杆菌质粒的集合中共表达蛋白质对。串联质粒以不依赖连接的方式形成，有效繁殖，并大量产生蛋白质对。这极大地促进了结构基因组学项目的共表达，该项目产生了数千个具有相同来源和抗生素标记的克隆。
We developed a method to co-express protein pairs from collections of otherwise identical Escherichia coli plasmids expressing different ORFs by incorporating a 61-nucleotide sequence ( LINK) into the plasmid to allow generation of tandem plasmids. Tandem plasmids are formed in a ligation-independent manner, propagate efficiently, and produce protein pairs in high quantities. This greatly facilitates co-expression for structural genomics projects that produce thousands of clones bearing identical origins and antibiotic markers.