title: High throughput construction and small scale expression screening of multi-tag vectors in Escherichia coli.
abstract: A suite of protein fusion vectors is presented that has been designed so that nine separate fusion vectors can be constructed from one PCR product using InFusion™ cloning. These vectors in combination with a small scale Escherichia coli expression screen can be used to assess in parallel the effect of fusion tags on solubility. The vectors were tested with 20 target proteins and the results suggest that the vectors are useful both as a rescue strategy if the N-terminal hexa-histidine tagged construct does not express and also as part of a primary expression experiment.
authors.affiliation: Division of Structural Biology, Henry Wellcome Building for Genomic Medicine, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK. louise@strubi.ox.ac.uk
authors.forename: Louise E
authors.initials: LE
authors.lastname: Bird
mesh_terms: D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D057166:High-Throughput Screening Assays; D006639:Histidine / Q000378:metabolism; D006801:Humans; D051379:Mice; D008967:Molecular Biology / Q000379:methods*; D008969:Molecular Sequence Data; D009345:Neisseria meningitidis; D009842:Oligopeptides / Q000378:metabolism; D011550:Pseudomonas aeruginosa; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D012995:Solubility
chemical_list: C471213:His-His-His-His-His-His; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; D006639:Histidine