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Fusion protein of CTB (Cellulose Tribenzoate), human insulin and glutamic acid decarboxylase 3p531 fragments and application thereof

A glutamic acid decarboxylase and fusion protein technology, applied in the fields of application, peptide/protein components, lyase, etc., to achieve high yield, stable gene expression products, and high activity

Active Publication Date: 2014-05-28
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Most of the drugs for treating diabetes in the prior art are aimed at type 2 diabetes, and there are relatively few drugs for treating type 1 diabetes

Method used

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  • Fusion protein of CTB (Cellulose Tribenzoate), human insulin and glutamic acid decarboxylase 3p531 fragments and application thereof
  • Fusion protein of CTB (Cellulose Tribenzoate), human insulin and glutamic acid decarboxylase 3p531 fragments and application thereof
  • Fusion protein of CTB (Cellulose Tribenzoate), human insulin and glutamic acid decarboxylase 3p531 fragments and application thereof

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Experimental program
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Embodiment 1

[0032] 1. Construction of fusion gene

[0033] (1) Use the pBac-CTB-INS plasmid as a template and aF / aR as primers to amplify to obtain the target fragment CTB-INS-GPGP;

[0034] Upstream primer aF: 5′-CG GGATCC ATGATTAAATTAAAATTTGG-3';

[0035] Downstream primer aR: 5′-GGGGCCGGGGCCGTTGCAGT

[0036] AGTTCTCCAGCTGGTAGAGG-3';

[0037] The reaction conditions are: pre-denaturation at 94°C for 5 minutes, amplification at 94°C for 30 seconds, annealing at 60°C for 30 seconds, extension at 72°C for 1 minute, 30 cycles of reaction, and finally extension at 72°C for 10 minutes;

[0038] (2) Use the pBac-CTB-3p531 plasmid as a template and bF / bR as primers to amplify to obtain the target fragment GPGP-3p531;

[0039] Upstream primer bF: 5′-GGCCCCGGCCCCCCGGTTATCAAAAGCTCG-3′;

[0040] Downstream primer bR: 5′-GG GAATTC TTAAACCATGGTAGTACCGTA-3';

[0041] The reaction conditions are: pre-denaturation at 94°C for 5 minutes, amplification at 94°C for 30 seconds, anneal...

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Abstract

The invention discloses a fusion protein of CTB (Cellulose Tribenzoate), human insulin and glutamic acid decarboxylase 3p531 fragments and application of the fusion protein. The application of the fusion protein is in preparing a medicament for treating type I diabetes. Silkworm larvae or silkworm chrysalides infected by recombinant silkworm baculoviruses efficiently express the fusion protein which has higher activity than that of the fusion protein expressed by prokaryotes directly and higher yield than that of the fusion protein expressed by transgenic plants; a silkworm body has multiple natural protein protective agents, thereby having a protection function for expression products and facilitating the more stable expression products; the prepared fusion protein can be prepared into the medicament for treating the type I diabetes and relieving injection pains and infection threats for patients.

Description

technical field [0001] The invention relates to the technical field of biopharmaceutical development, in particular to a fusion protein of CTB, human insulin and glutamic acid decarboxylase 3p531 fragment and application thereof. Background technique [0002] Diabetes mellitus is a metabolic disease characterized by high blood sugar caused by insulin secretion defect or insulin action disorder. It has become one of the major diseases affecting human health in the world. Diabetes is divided into type Ⅰ and type Ⅱ. Among them, type Ⅰ diabetes (insulin-dependent diabetes, IDDM) is an autoimmune disease caused by the destruction of pancreatic β cells, resulting in defective insulin secretion, while the ability of insulin secretion in patients with type Ⅱ diabetes is not It is not completely lost, and some patients even produce too much insulin in the body, but the effect of insulin is poor, resulting in a relative lack of insulin in the body. [0003] Type 1 diabetes accounts f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/60C12N7/01C12N9/88A61K38/51A61P3/10C12R1/93
CPCY02A50/30
Inventor 金勇丰刘宝平杨赟周陈娟岳媛潘华伟李司
Owner ZHEJIANG UNIV
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