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

被引:89
作者
Alexandrov, A
Vignali, M
LaCount, DJ
Quartley, E
de Vries, C
De Rosa, D
Babulski, J
Mitchell, SF
Schoenfeld, LW
Fields, S
Hol, WG
Dumont, ME
Phizicky, EM
Grayhack, EJ [1 ]
机构
[1] Univ Rochester, Med Ctr, Ctr Human Genet & Mol Pediat Dis, Rochester, NY 14642 USA
[2] Univ Rochester, Med Ctr, Struct Genom Pathogen Protozoa Consortium, Rochester, NY 14642 USA
[3] Univ Rochester, Med Ctr, Dept Biochem & Biophys, Rochester, NY 14642 USA
[4] Univ Washington, Dept Genome Sci, Seattle, WA 98195 USA
[5] Univ Washington, Dept Med, Seattle, WA 98195 USA
[6] Univ Washington, Howard Hughes Med Inst, Seattle, WA 98195 USA
[7] Univ Washington, Dept Biochem, Seattle, WA 98195 USA
关键词
D O I
10.1074/mcp.T400008-MCP200
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:934 / 938
页数:5
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