Method for preparing chicken akirin2 protein using insect cell expression system

Abstract

The invention discloses a method of preparing chicken Akirin2 protein through an insect cell expression system. The method includes the steps of: 1) performing codon optimization to the chicken Akirin 2 gene according to insect cell preferred codons; 2) synthesizing an Akirin2 sequence; 3) PCR-amplifying the Akirin2; 4) constructing a baculovirus expression carrier pFastBac1-his-thrombin-Akirin2; 5) performing pFastBac1-his-thrombin-Akirin2 baculovirus carrier package in the insect cell sf9; and 6) expressing and purifying the chicken Akirin2 virus protein. The method has high expression level and can be used for producing the protein having proper translation, which is convenient to modify the protein, thereby simplifying cell growth conditions.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a method for preparing Akirin2 protein using an insect cell expression system. Background technique [0002] Akirin is an innate immune protein discovered by Goto et al. in 2008 when they conducted a functional genome-wide RNAi screen on Drosophila cells. Akirin consists of 201 amino acids and is highly conserved among different species. Two genes, Akirin1 and Akirin2, have been found in higher mammals including humans, cattle, mice, rats and dogs, and the homology of Akirin2 among mammals is significantly higher than that of Akirin1. In chicken there is only the akirin2 gene, which is not found in plants, bacteria and yeast. [0003] Studies have shown that Akirin2 plays an important role in the immune inflammatory response of Drosophila and mice. It is an essential nuclear factor for innate immune response signaling pathways such as Imd and TLR, and is involved in NF-κB-dep...

AI Technical Summary

Problems solved by technology

[0004] Commonly used protein expression systems mainly include prokaryotic expression systems, which have high protein expression levels, but the products are often inclusion bodies, which do not have biological activity; yeast expression systems have incomplete glycosylation modifications for expressed products; mammalian cell expression systems Although the expression product can be modified by glycosylation to produce an active protein, its expression level is low, it is not easy to amplify, and the cell growth conditions are relatively high

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