Process for producing human insulin using transgenic tomato

Abstract

The present invention utilizes transgenic tomato as bioreactor to produce human insulin. The human insulin is produced through designing plant preference codon, artificially synthesizing several DNAsegment, splicing, and adding endoplasmic reticulum residence sequence of KDEL in the C terminal to avoid the degradation of insulin in plant cell. The gene is transformed into tomato via agrobacterium mediating process under the driving of CaMV35S promoter and fruit specificity promoter 2A12 to express human insulin in tomato fruit. This kind of tomato capable of producing human insulin may become one kind of convenient oral product for preventing and treating some diabetes of depending insulin and autoimmune dysfunction, and may be used to extract human insulin for making injection.

Description

Technical field: [0001] The invention relates to a method for producing human insulin by transgenic plants and its product. Background technique: [0002] Bioreactors are an area of ​​great concern to many biotechnology R&D departments in the near future. Human growth factors are produced in mammary bioreactors such as dairy cows and dairy goats. In the research of plant bioreactor, the research of using potato tubers to produce hepatitis B vaccine has received extensive attention. Lettuce and tomato, as vegetables or fruits that can be eaten raw, have great application potential in the production of oral polypeptide drugs. Human insulin is very important for the treatment of insulin-dependent diabetic patients. Metabolic regulation in some diabetic patients is associated with autoimmune function. [0003] Oral insulin has been shown to be effective in the treatment of certain types of diabetes [Reddy S et at, Pancreas 2000, 20(1): 55-60; Ploix C et at, Diabetes 1999, 48...

AI Technical Summary

Problems solved by technology

At the same time, the dendritic cells scattered in the intestinal cells can also take up the antigens in the intestinal lumen and activate the immune response through the circulation of the lymphatic system, but whether this process induces tolerance or immunity is still unclear

[0006] The treatment of oral tolerance has a certain therapeutic effect on the disease because it can elicit an antigen-specific immune response, but it is limited by some practical problems, that is, this therapy requires a large amount of protein in both humans and experimental animals antigen

To address this issue, several traditional expression systems, including microbial fermentation and mammalian cells, have been experimented with, however, they all have methodological limitations.

A major problem with bacterial expression systems is their inability to perform the post-translational processing necessary for many mammalian proteins, such as glycosylation

While the mammalian cell expression system overcomes this problem, its expensive culture equipment and its relatively low yield make it expensive and impractical

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