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Expresso® Cloning and Expression Systems streamline recombinant protein expression
Author(s) -
Sen Saurabh,
Sternard Heather,
Franz Laura,
Drinkwater Colleen,
Sheets Lynne,
Niebauer Chris,
Mead David,
Godiska Ron,
Steinmetz Eric J
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.584.1
Subject(s) - cloning (programming) , multiple cloning site , expression vector , recombinant dna , biology , lac operon , cloning vector , promoter , microbiology and biotechnology , target protein , vector (molecular biology) , gene , molecular cloning , computational biology , gene expression , genetics , computer science , programming language
The Expresso® Cloning and Expression Systems use Expressioneering™ Technology, an in vivo recombinational cloning strategy, to dramatically increase the speed, efficiency and success of target gene cloning and soluble protein expression in E. coli and mammalian system. Expressioneering allows directional cloning of PCR products into pre‐processed expression vectors without sample clean‐up or enzyme treatment. Expresso System vectors were constructed with various inducible promoters, N‐ or C‐terminal 6xHis tags, and an optional cleavable SUMO tag for improved recombinant protein solubility in bacterial systems. The Expresso T7 System uses an IPTG‐inducible T7‐lac promoter and the Expresso Rhamnose Systems features the tunable, rhamnose‐inducible, rhaP BAD promoter, and uses a single host strain for both cloning and expression. The new Expresso CMV system for mammalian protein production utilizes the same Expressioneering technology to provide rapid, precise, and directional insertion of fragments into the cloning site. The small size of the pME‐HA vector (3.4 kb) and minimal amount of bacterial DNA sequence supports robust protein expression, higher transfection efficiency and simplifies subsequent manipulations. The Expresso CMV vector allows addition of the HA tag for immono‐detection or affinity purification.