Abstract
Human annexin V cDNA was cloned into plasmid pET19b and fused to a ten consecutive histidine tag at N-terminal. When expressed in E. coli BL21(DE3) LysS, the recombinant His10-annexin V accumulated in soluble form in the cytoplasm. By two-step chromatography, i.e., metal chelate affinity chromatography and anion exchange chromatography, recombinant His10-annexin V was purified to homogeneity on silver-stained SDS-PAGE gel. Recombinant annexin V, 7.4 mg, was obtained from a 1 litre flask culture.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Annexin A5* / genetics
-
Annexin A5* / isolation & purification
-
Annexin A5* / metabolism
-
Cloning, Molecular
-
DNA, Complementary
-
Electrophoresis, Polyacrylamide Gel
-
Enzyme-Linked Immunosorbent Assay
-
Escherichia coli / genetics*
-
Escherichia coli / metabolism
-
Humans
-
Plasmids / genetics*
-
Plasmids / metabolism
-
Recombinant Fusion Proteins / genetics
-
Recombinant Fusion Proteins / isolation & purification
-
Recombinant Fusion Proteins / metabolism
Substances
-
Annexin A5
-
DNA, Complementary
-
Recombinant Fusion Proteins