Induced expression of fusion protein taking TTC as neuronal cell targeting gene

Abstract

The invention provides induced expression of a fusion protein taking TTC as a neuronal cell targeting gene, and belongs to the technical field of biological medicines. The method comprises the following steps: 1, respectively designing corresponding primers for base sequences of mitochondrial outer membrane protein FUNDC1, tetanus heavy chain C-terminal non-toxic fragment TTC and green fluorescent protein EGFP, and then respectively carrying out PCR (Polymerase Chain Reaction) amplification; 2, recovering the genes of the three segments in the step 1, connecting the genes with a PET-32a (+) expression vector carrying an HIS (6 * his) label, and then transforming escherichia coli DH5alpha; 3, extracting plasmids for sequencing through positive clone screening; 4, transferring the plasmids which are correctly sequenced into escherichia coli BL21 (DE3), and carrying out mass expression and purification, so as to obtain the TTC-mFunddc1-EGFP fusion protein with relatively high concentration; the neuroinvasiveness of a tetanus non-toxic C-terminal fragment TTC in the TTC-mFunddc1-EGFP fusion protein is utilized, and the tetanus non-toxic C-terminal fragment TTC is targeted to nerve cells. According to the invention, the mouse FUNDC1 fusion protein with targeted nerve cells can be obtained.

Description

technical field [0001] The invention belongs to the technical field of biomedicine and relates to the induced expression of a fusion protein using TTC as a neuron cell targeting gene. Background technique [0002] Diabetes mellitus (DM) is a major global public health problem. The incidence of diabetes is increasing year by year, and it has become the third largest non-communicable disease in developed countries after cardiovascular and tumors. According to epidemiological research surveys, the total number of diabetes patients in the world is about 425 million, and my country's diabetes patients account for about 1 / 3 of the world. With the improvement of living standards and changes in lifestyle, the incidence of diabetes is increasing. Diabetic neuropathy is an important complication of diabetes, and it has various forms, mainly including peripheral neuropathy and autonomic neuropathy, etc. These lesions seriously affect the quality of life of diabetic patients, and oxida...

AI Technical Summary

Problems solved by technology

[0008] 1. The huge genome of HSV-1 makes it difficult to construct recombinant viral vectors by conventional methods

[0009] 2. This method is cumbersome to operate, low in efficiency of recombination, time-consuming and labor-intensive

[0010] 3. However, its potential risks are still to be determined. HSV has neurocytotoxic effects and immune responses, which can cause more obvious local inflammation and necrosis

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