Escherichia coli expression vectors containing a protein kinase recognition motif, His6-tag and hemagglutinin epitope.
Escherichia coli expression vectors, based on the pET system, were constructed to allow fusion of a protein kinase (PK) recognition motif, a hemagglutinin (HG) epitope-tag and a His6-tag at the N-terminal portion of a protein of interest. The fusion proteins, that result from expression using these vectors, can be phosphorylated in vitro using cAMP-dependent PK, immunoprecipitated using monoclonal antibody against the HG-epitope, and can be rapidly purified using a Ni2+ column.
Kelman
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Z
Microbiology Department Hearst Research Foundation, Cornell University Medical College, New York, NY 10021, USA.
Yao
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N
O'Donnell
M
M
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D000939:Epitopes; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D006388:Hemagglutinins / Q000276:immunology; D006639:Histidine; D008969:Molecular Sequence Data; D011494:Protein Kinases / Q000378:metabolism; D011993:Recombinant Fusion Proteins
D000939:Epitopes; D006388:Hemagglutinins; D011993:Recombinant Fusion Proteins; D006639:Histidine; D011494:Protein Kinases