Method for high-efficiency expression of all subtype mature proteins of IL-37 by utilizing tobaccos

Abstract

The invention discloses a method for high-efficiency expression of all subtype mature proteins of IL-37 by utilizing tobaccos. The method comprises the following steps of: inserting coding sequences of TEV protease and enzyme cutting sites thereof between coding sequences of signal peptide and mature IL-37 proteins to form an IL-37-TEV fusion gene; according to the preference of tobacco codons, under the condition of guaranteeing no change of the amino acid sequence, optimizing the sequence of the fusion gene; connecting 5'-end upstream of the constructed IL-37-TEV fusion gene with a common promoter including other regulating areas, and connecting the 3'-end downstream with a transcription terminator; integrating a constructed recombinant expression vector onto cell chromosomes of the tobacco to culture a transgenic tobacco plant with high-efficiency stable expression of IL-37 and injecting the constructed recombinant expression vector to the tobacco leaves to instantaneously express target proteins. The method disclosed by the invention has the advantages that by construction of the plant expression vector, the mature proteins of IL-37 with high activity are expressed in the plants, and the obtained product has important application value for treatment of numerous inflammatory diseases and early detection and diagnosis of the inflammatory diseases.

Description

technical field [0001] The invention relates to the fields of genetic engineering and protein expression, in particular to a method for efficiently expressing mature proteins of IL-37 subtypes by using tobacco. Background technique [0002] Human interleukin-37 (IL-37) is a newly added member of the IL-1 family of cytokines, discovered in 2000. In contrast to other members of the IL-1 family, IL-37 is an anti-inflammatory cytokine with broad anti-inflammatory effects, playing both a role in limiting innate inflammation and suppressing acquired immune responses. plays a key role. In vitro experiments have found that inhibiting the normal expression of IL-37 in human cells will increase the expression and release of inflammatory cytokines induced by IL-1, Toll-link receptor (TRL) and uric acid crystals, including TNF-α, IL-1α, IL-1ß, IL-6 and G-CSF. Human IL-37 is active in mice, although no human equivalent IL-37 gene has been identified in mice so far. The constructed IL...

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