Human profibrinolysin kringle 5 deletion mutation recombinant polypeptide
A human plasminogen, recombinant polypeptide technology, applied in the direction of recombinant DNA technology, enzymes, enzymes, etc., can solve the problems of vascular leakage without satisfactory treatment methods, etc.
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[0016] Materials: Retinal capillary endothelial cells (RCEC) and epithelial cells were isolated from bovine eyes as described previously (Grant, M.B. et al. Invest Ophthalmol. Vis. Sci., 32:53-64, 1991). It is shaped like a pebble, bound to fluorescent probe DiI-labeled acetylated low-density lipoprotein (Biomedical Technologies Inc., Stoughton, MA), confirmed as RCEC, cultured epithelial cell purity by α-smooth muscle actin-specific FITC binding Antibody identification. Brown-Norway rats were purchased from Harlan (Indianapolis, IN).
[0017] 1. Construction of K5mut1:
[0018]Human K5 cDNA was amplified from liver RNA by RT-PCR (Ma J.X. et al. FEBS Lett, 452:199-204, 1999). The deletion mutant was amplified by PCR using the human K5 template. The 5'PCR primer (5'-ATGAATTCGTGTATGTTTGGGAATGGG-3') and 3'PCR primer (5'-GCCAAGCTTACACTGAGGGACATCACAGTAG-3') each contained EcoR I and HindIII sites to facilitate cloning. PCR deleted the nucleotide sequence encoding 10 amino acid ...
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