Preparation method of fused protein for antioxidation and preventing diabetes and application thereof

Abstract

The invention discloses a making method and application of diabetes-proof fusion protein, which is characterized by the following: designing up-stream and down-stream primers P1 and P2 according to the gene sequence of ProT alpha; guiding BamHI enzyme cutting point in the P1 and EcoRI enzyme cutting point in the P2; using RT-PCR method to obtain the original full-length gene cDNA of prothymosin alpha; purifying the PCR product to connect with pMD18-T Vector; inverting DH5 alpha competence cell; comparing 7 monoclonal gene sequences and reference proT alpha gene sequences; cutting the product of ProT alpha PCR through EcoRI and BamHI enzyme; plugging ProT alpha gene segment into prokaryotic expressive carrier pGEX 6P-1 cut by the same enzymes; inverting escherichia coli BL21; adding IPTG; inducing the expression; centrifuging bacteria liquid; dissolving the sediment through sampling buffer; centrifuging to fetch the supernatant to do electrophoresis; obtaining the GST-Pro alpha fusion protein.

Description

technical field [0001] The present invention relates to a fusion protein, in particular to a preparation method and application of a fusion protein for anti-oxidation and prevention of diabetes. Background technique [0002] Prothymosin alpha (prothymosin alpha) is a small acidic protein with a molecular weight of 13,000, originally isolated from the rat thymus. Although thymosin is secreted by the thymus, prothymosin alpha is expressed in many tissues in vivo and is a conserved DNA sequence without introns. The clinical application of thymosin began in the mid-1970s, and its most important contribution is the treatment of thymus-dependent diseases. The activity of sex lymphocytes plays a major role, so it is widely used in the treatment of systemic lupus erythematosus, Behçet's syndrome, rheumatoid arthritis, severe infection, bronchial asthma, acute and chronic hepatitis and tumors. In addition, because human aging obviously reflects the atrophy and functional failure of...

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Problems solved by technology

Induced mouse thymus atrophy, kidney enlargement, and serum antioxidant capacity decreased significantly

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