[The construction and expression of two humanized scFv-urokinase fusion genes].
By using PCR and DNA recombination, two fusion genes of humanized mouse anti-human fibrin scFv and low molecular weight single chain urokinase (Scu-PA-32K) was constructed. The difference of these two fusion genes lay in the linker between two moieties, one was (Ala)3 and another was (Gly4Ser)3. These two fusion genes were both overexpressed in E. coli with the expression level at 30%. Both expression products showed the activity of binding antigen D-Dimer and activating plasminogen after the denaturation and renaturation, but under general refolding conditions, the one with linker (Gly4Ser)3 showed better effect in the renaturation of fusion protein.
Liu
Z G
ZG
Instutute of Biotechnology, Academy of Military Medical Science, Beijing.
Yu
W Y
WY
Wang
X
X
D000818:Animals; D015870:Gene Expression; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics*; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D014568:Urokinase-Type Plasminogen Activator / Q000235:genetics*
D007128:Immunoglobulin Fragments; D011993:Recombinant Fusion Proteins; C055966:immunoglobulin Fv; D014568:Urokinase-Type Plasminogen Activator