A novel peptide tag for detection and purification of recombinant expressed proteins.
Peptide tags have proven useful for the detection and purification of recombinant proteins. However cross reactions of antibodies raised to the tag are frequently observed due to the presence of host proteins containing all or parts of the tag. In this report we have identified a unique viral peptide sequence, R-tag, that by blast searches is absent from the commonly expression hosts Arabidopsis thaliana, Escherichia coli, Pichia pastoris and mouse myeloma cell NSO. We have prepared monoclonal antibodies to this peptide and confirmed the absence of this peptide sequence from the above genomes by Western blotting. We have also modified protein expression vectors to incorporate this sequence as a fusion tag in expressed proteins and shown its use to successfully purify recombinant proteins by immunoaffinity procedures.
Jones
William T
WT
The Horticulture and Food Research Institute of New Zealand Ltd, Private Bag 11 030, Palmerston North, New Zealand. wjones@hortresearch.co.nz
Harvey
Dawn
D
Kirk
Christopher
C
Rakonjac
Jasna
J
Sun
Xiaolin
X
Frearson
Nicky
N
Samarrai
Taha Al
TA
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000915:Antibody Affinity; D017360:Arabidopsis / Q000235:genetics / Q000276:immunology; D001483:Base Sequence; D002460:Cell Line; D003429:Cross Reactions; D003460:Crystallization; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000276:immunology; D005260:Female; D005822:Genetic Vectors; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008811:Mice, Inbred DBA; D009842:Oligopeptides / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D010843:Pichia / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D017931:DNA Primers; D009842:Oligopeptides; D011994:Recombinant Proteins