Chlamydomonas reinhardtii Se1K expression vector as well as construction method and expression method thereof

Abstract

The invention provides a construction method of a chlamydomonas reinhardtii Se1K expression vector. The construction method comprises the following steps: amplifying an ORF gene segment of human Se1K to obtain an ORF gene amplification product; amplifying an SECIS gene segment of chlamydomonas reinhardtii Se1W1 to obtain an SECIS gene amplification product; recombining the ORF gene amplification product with the SECIS gene amplification product to obtain a recombinant Se1K gene; and introducing the recombinant Se1K gene into a plasmid vector which can be expressed in the chlamydomonas reinhardtii to form a recombinant plasmid product. According to the chlamydomonas reinhardtii Se1K expression vector constructed by the method, a novel plasmid vector capable of expressing an exogenous target selenoprotein K in the chlamydomonas reinhardtii is constructed; compared with the existing in-vitro expression vectors such as a prokaryotic expression system, and a method, the product is generated by a similar expression mechanism, so that the product has good monotonicity and directionality; the efficiency is greatly improved; the chlamydomonas reinhardtii can be adopted as a reactor for producing selenoprotein during expression; and the construction method is portable and efficient, and can completely meet the bases of research on structure functions of the selenoprotein and development of selenoprotein anti-cancer drugs.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a Chlamydomonas reinhardtii SelK expression vector, a construction method and an expression method thereof. Background technique [0002] As an essential trace element for humans and animals, selenium plays an important role in many physiological functions and is closely related to many diseases, such as cancer, neurodegenerative diseases and cardiovascular diseases. Selenium mainly exerts biological functions through various selenoproteins. For example, among the selenoproteins discovered so far, SelK selenoprotein participates in coping with oxidative stress in cardiomyocytes and in regulating endoplasmic reticulum stress in human liver cancer cells. Promote calcium flow in the process of immune cell activation mediated by body; participate in the degradation of endoplasmic reticulum-related proteins, participate in the regulation of CD36 palmitoylation ...

AI Technical Summary

Problems solved by technology

However, the production efficiency of selenoproteins is very low, and it is impossible to achieve large-scale expression of selenoproteins, nor can it be purified to protein

Therefore, the in vitro expression and synthesis of selenoproteins cannot meet the single, efficient, and large-scale application requirements.

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