A kind of preparation method of modified polyester material
A polyester, modified technology, applied in the field of polyester material preparation, to prevent thrombus blockage, reduce injury, facilitate tissue healing and anti-coagulation effects
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Embodiment 1
[0019] 1. Transfect Escherichia coli (BL21) cells with the prokaryotic system expression vector carrying the first polypeptide F1 gene, coat it on Luria-Bertani solid medium containing ampicillin, put it upside down and put it in a biochemical incubator at 37°C for 14 ~16 hours; pick a single colony and put it into 4mL Luria-Bertani liquid medium containing fresh ampicillin, insert it into an air shaker with a shaking speed of 200r / min at 37°C for 8-10 hours; Amplify and culture in 1L fresh Luria-Bertani liquid medium containing ampicillin at a ratio of 1:100; when the density of the bacterial solution reaches OD 600 When = 0.3-1.8, 0-1.2 mM isopropyl-β-D-thiogalactoside was added to induce culture for 1-8 hours, and the bacterial cells were collected by centrifugation at 4°C.
[0020] 2. Resuspend and mix the collected bacterial cells with the binding buffer of glutathione transferase (GST) affinity chromatography, and put them on ice to ultrasonically disrupt the cells and r...
Embodiment 2
[0026] 1. Transfect Escherichia coli (BL21) cells with the prokaryotic system expression vector carrying the first polypeptide F1 gene, coat it on Luria-Bertani solid medium containing ampicillin, put it upside down and put it in a biochemical incubator at 37°C for 14 ~16 hours; pick a single colony and put it into 4mL Luria-Bertani liquid medium containing fresh ampicillin, insert it into an air shaker with a shaking speed of 200r / min at 37°C for 8-10 hours; Amplify and culture in 1L fresh Luria-Bertani liquid medium containing ampicillin at a ratio of 1:100; when the density of the bacterial solution reaches OD 600 When =0.3-1.8, add 0-1.2mM isopropyl-β-D-thiogalactoside to induce culture for 1-8 hours, and collect bacterial cells by centrifugation at 4°C.
[0027]2. Resuspend and mix the collected bacterial cells with the binding buffer of glutathione transferase (GST) affinity chromatography, put them on ice and ultrasonically break the cells to release the protein; after ...
Embodiment 3
[0033] 1. Transfect Escherichia coli (BL21) cells with the prokaryotic system expression vector carrying the second polypeptide F4 gene, coat it on the Luria-Bertani solid medium containing ampicillin, and place it upside down in a biochemical incubator at 37°C for 14 ~16 hours; pick a single colony and put it into 4mL Luria-Bertani liquid medium containing fresh ampicillin, insert it into an air shaker with a shaking speed of 200r / min at 37°C for 8-10 hours; Amplify and culture in 1L fresh Luria-Bertani liquid medium containing ampicillin at a ratio of 1:100; when the density of the bacterial solution reaches OD 600 When = 0.3-1.8, 0-1.2 mM isopropyl-β-D-thiogalactoside was added to induce culture for 1-8 hours, and the bacterial cells were collected by centrifugation at 4°C.
[0034] 2. Resuspend and mix the collected bacterial cells with the binding buffer of glutathione transferase (GST) affinity chromatography, put them on ice and ultrasonically break the cells to release...
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