Human Scurfin protein span with membrane penetration sequence, fragment and preparation method

Abstract

The invention relates to the technical fields of gene engineering and the biomedicine of protein transduction, in particular to fusion proteins of a human Scurfin span with a membrane penetration sequence, a Forkhead removing domain human Scurfin fragment and a proline-rich removing domain human Scurfin fragment and a preparation method. The fusion proteins of the human Scurfin span with the membrane penetration sequence, the Forkhead removing domain human Scurfin fragment and the proline-rich removing domain human Scurfin fragment respectively have amino acid sequences shown in SEQ ID NO.1, SEQ ID NO.2 and SEQ ID NO.3. The invention adopts a mode of constructing PTD-hScurfin to obtain the fusion proteins for the first time, transduces a target protein into Jurkat T and peripheral CD4+CD25-T cells by the direct mediation and fusion of a protein membrane penetration structure field to obtain Treg in mass, and breaks through the bottleneck constraining Treg application in the hope of being applied to the repulsion-resisting treatment of clinical organ transplantation finally.

Description

technical field [0001] The invention relates to the biomedical technical field of genetic engineering and protein transduction. Specifically, the present invention relates to the full-length human Scurfin with a membrane-penetrating sequence, the human Scurfin fragment with the forkhead domain (ΔForkhead domain, ΔFKH) removed, and the human Scurfin fragment with the proline-rich domain (ΔProline-rich domain, ΔPRD) removed. The fusion protein of Scurfin fragment: the recombinant construction of PTD-hScurfin, PTD-ΔhFKH and PTD-ΔhPRD gene sequence and the fusion protein prepared therefrom has the effect on Jurkat T cells (human acute lymphoblastic leukemia cell line) and human peripheral blood CD4 + The role of T cells in inhibiting proliferation, regulating cytokine secretion and inducing immune tolerance, as well as the application of regulatory T-like cells in the preparation, prevention and treatment of inflammatory immune-related diseases (including autoimmune diseases, tran...

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Problems solved by technology

[0021] (3) Impact on transplant rejection

[0039] The application of Treg in the induction of transplant immune tolerance has begun at home and abroad, but the number of natural Treg is small and difficult to expand. Therefore, in recent years, many laboratories have begun to use T cells with regulatory functions after Foxp3 gene transfection to induce transplant immunity. Tolerance, and achieved success in animal experiments, but the biggest disadvantage of transgenics is low transfection efficiency (except for retroviral vectors), difficult expression regulation, and potential danger, especially retroviral vectors can induce gene mutations The possibility. In 2005, Veldhoen et al. used enzyme-deficient Lck-PTD to transduce initial CD4 + T cells have successfully induced T cells to acquire regulatory functions. At present, there are no reports at home and abroad that use PTD and Foxp3 genes to construct and express fusion proteins for basic and clinical research.

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