Preparation method for novel fusion protein reversing renal fibrosis and activity detection

Abstract

The invention belongs to the field of medicinal biotechnology, in particular relates to a novel fusion transmembrane protein capable of cloning genes, expressing exogenous genes in prokaryotic cells, purifying objective protein, and inhabiting TGF-beta1 passages to reverse renal fibrosis. The novel fusion transmembrane protein is characterized in that a TGF-beta1 passage is used for inhabiting protein SARA which is fused with a transmembrane protein domain (PTD) in colon bacillus to obtain efficient expression, an ammonium sulfate salting out method and an anion-cation exchange method are utilized to purify objective protein, and the activity of the obtained objective protein is tested. The invention has the advantage of high efficiency and low toxicity of a novel TGF-beta1 passage inhibitor SARA, and can effectively pass through epicyte and karyotheca, thereby reducing the clinical applied dosage of the TGF-beta1 passage inhibitor and enhancing the function of the inhibitor in treatment of reversing renal fibrosis. A novel peptide drug can be expected to obtain for clinical treatment of reversing renal fibrosis.

Description

technical field [0001] The invention belongs to the field of medical biotechnology, and specifically relates to gene cloning, expression of exogenous genes in prokaryotic cells, preparation method and activity detection of target protein, especially preparation method and activity detection of a novel fusion protein for reversing renal fibrosis. Background technique [0002] (1) Reversal of renal fibrosis treatment related research and existing problems: [0003] Renal fibrosis is the ultimate common outcome of chronic kidney disease caused by different etiologies (including inflammation, injury, drugs, diabetes and genetic factors, aging, etc.). Various progressive kidney diseases, regardless of the primary disease, will eventually lead to renal fibrosis, progress to end-stage renal failure, and must rely on dialysis or kidney transplantation to maintain life. Therefore, studying the pathogenesis of renal fibrosis, exploring effective treatments that can alleviate the proc...

AI Technical Summary

Problems solved by technology

However, the current chemical drugs or gene therapy methods that block the TGF-β1 signal transduction pathway are difficult to be clinically accepted due to safety issues and the possibility of aggravating renal inflammatory responses.

The activation of the BMP7 signal transduction pathway requires a very high dose of BMP7 and is expensive, making it difficult to use recombinant BMP7 in the treatment of renal fibrosis in clinical practice.

[0019] At present, there is no research report on the direct use of recombinant SARA peptide aptamer to prevent and treat renal fibrosis. Therefore, by preparing recombinant SARA peptide aptamer and studying its therapeutic effect on renal fibrosis, we explore the possibility of replacing BMP7 and effectively treating, Protein Drugs to Alleviate Renal Fibrosis

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