Preparation and application of thrombolytic enzyme
A technology of thrombolytic enzyme and fibrinolysis, applied in the field of thrombolytic enzyme, can solve problems such as being in the experimental stage, and achieve the effects of high-efficiency dissolution, significant effect, and blood supply recovery.
Inactive Publication Date: 2020-09-22
SICHUAN ACAD OF CHINESE MEDICINE SCI
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Problems solved by technology
Such drugs have been improved and improved in terms of specificity, half-life, and thrombolytic efficiency, but most of them are currently in the experimental stage
Method used
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Embodiment Construction
[0026] The present invention will be further described according to the following examples, and the mode of the present invention includes but not limited to the following examples.
[0027] The strains, vectors, and media used in this example are Escherichia coli Trelief 5a-pClone007 and BL21(DE3)-pDE1 for cloning and overexpression respectively. Escherichia coli was cultured in LB medium with shaking at 37°C. LB selection medium added with 50 μg / ml kana is used for screening, cultivating recombinant Escherichia coli and producing recombinant protein.
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The invention discloses preparation and application of a thrombolytic enzyme. Bacillus velezensis with fibrinolytic activity is applied, a polymerase chain reaction technology is adopted to amplify and extract genome DNA, an AprE9912D gene is inserted into a pDE1 expression vector, and IPTG is utilized to induce escherichia coli to express a crude protein product. Through a preparation and purification process, the thrombolytic enzyme is obtained. The thrombolytic enzyme disclosed by the invention can specifically and efficiently dissolve thrombus, rapidly recover blood supply of embolism tissues and improve functions of the embolism tissues, is used for preventing and treating vascular embolism diseases, particularly cardiovascular and cerebrovascular embolism diseases, and has a remarkable effect, the efficiency of obtaining a target product is high, and the activity of the target product is higher than that of existing thrombolytic enzyme preparations.
Description
technical field [0001] The invention relates to the field of thrombolytic enzymes, in particular to the preparation and application of a thrombolytic enzyme. Background technique [0002] According to epidemiological studies, the incidence and level of risk factors for cardiovascular and cerebrovascular diseases in my country are on the rise. Thrombosis is the main cause of cardiovascular disease, including arterial thrombosis, venous thrombosis and microcirculation thrombosis. Thrombotic diseases are the main cause of cardiovascular and cerebrovascular diseases, and are also the main factors leading to death or disability. Seeking thrombolytic drugs for clinical treatment has important medical value. In the treatment of thrombotic diseases, plasmin has potential application prospects. [0003] At present, thrombolytic drugs have been developed to the fourth generation. The first generation is represented by streptokinase (SK) and urokinase (UK), which are characterized b...
Claims
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IPC IPC(8): C12N9/54C12N15/70A61K38/16A61P9/00A61P9/10A61P7/02C12R1/19
CPCA61K38/166A61P7/02A61P9/00A61P9/10C12N9/54C12N15/70
Inventor 谭正怀张秀
Owner SICHUAN ACAD OF CHINESE MEDICINE SCI
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