The human fibrinogen B beta chain was expressed in Escherichia coli to study the functions of fibrinogen associated with this subunit. Recombinant B beta chains were expressed at 100 ng/mL in an IPTG- dependent manner. A first cistron sequence, inserted into the expression vector 5′ to the B beta chain cDNA, was required to express the protein. Recombinant B beta chains were expressed within five minutes after induction with IPTG and were soluble in physiologic buffers. The recombinant B beta chains migrated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) at a rate identical to B beta chains from fibrinogen treated with N-glycanase. Recombinant B beta chains were cleaved by thrombin, as demonstrated by the loss of cross-reactivity with a monoclonal antibody (MoAb) specific for the undigested B beta 1–42 fragment. The levels of expression of the B beta chain were much lower than those reported previously for the gamma chain of fibrinogen expressed in a similar vector in E coli. However, these levels are sufficient to allow further characterization of this fibrinogen subunit.
ARTICLES| April 1, 1989
Expression in Escherichia coli of the human fibrinogen B beta chain and its cleavage by thrombin
Blood (1989) 73 (5): 1202-1206.
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MG Bolyard, ST Lord; Expression in Escherichia coli of the human fibrinogen B beta chain and its cleavage by thrombin. Blood 1989; 73 (5): 1202–1206. doi: https://doi.org/10.1182/blood.V73.5.1202.bloodjournal7351202
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