一种用于蛋白质对高通量共表达的简便方法。

A facile method for high-throughput co-expression of protein pairs.

作者信息

Alexandrov Andrei, Vignali Marissa, LaCount Douglas J, Quartley Erin, de Vries Christina, De Rosa Daniela, Babulski Julie, Mitchell Sarah F, Schoenfeld Lori W, Fields Stanley, Hol Wim G, Dumont Mark E, Phizicky Eric M, Grayhack Elizabeth J

机构信息

Structural Genomics of Pathogenic Protozoa (SGPP) Consortium and Center for Human Genetics and Molecular Pediatric Disease, University of Rochester Medical Center, Rochester, NY 14642, USA.

出版信息

Mol Cell Proteomics. 2004 Sep;3(9):934-8. doi: 10.1074/mcp.T400008-MCP200. Epub 2004 Jul 7.

DOI:10.1074/mcp.T400008-MCP200

PMID:15240823

Abstract

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.

摘要

我们开发了一种方法，通过将一段61个核苷酸的序列（LINK）整合到质粒中，从表达不同开放阅读框（ORF）的相同大肠杆菌质粒集合中共同表达蛋白质对，从而产生串联质粒。串联质粒以不依赖连接的方式形成，能高效繁殖，并大量产生蛋白质对。这极大地促进了结构基因组学项目中的共表达，该项目会产生数千个具有相同起源和抗生素标记的克隆。