Preparation method of mandarin fish rhabdovirus glycoprotein expressed by recombinant baculovirus

Abstract

The invention discloses a preparation method of mandarin fish rhabdovirus glycoprotein expressed by recombinant baculovirus. The preparation method comprises the following steps: designing and synthesizing primers; amplifying and purifying a target gene; constructing a recombinant donor plasmid pFastBac-G; constructing a recombinant shuttle plasmid rBacmid-G; preparing a recombinant baculovirus; purifying the target protein; and carrying out Western blot detection. According to the invention, a large amount of soluble recombinant proteins with good antigenicity and immunogenicity and similar functions to natural proteins can be obtained, and the method is suitable for high-efficiency expression of mandarin fish rhabdovirus glycoproteins and preparation of DNA vaccines.

Description

technical field [0001] The invention relates to a preparation method of mandarin fish rhabdovirus glycoprotein expressed by recombinant baculovirus, belonging to the field of biotechnology. Background technique [0002] Baculovirus expression system (BES) is currently one of the most widely used protein expression systems, commonly used in drug development, vaccines, growth-promoting factors, oncogenes, tumor suppressor gene protein products, and certain oncogenic virus proteins , immune active molecules, gene expression regulation and other fields. Using baculovirus as a carrier can efficiently express foreign genes. Up to now, hundreds of genes including animals, plants, viruses, bacteria, and fungi have been efficiently expressed in insect cells or larvae. The main feature of this expression system is that it can obtain a large number of soluble recombinant proteins with good antigenicity and immunogenicity and similar functions to natural proteins. After the recombinan...

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Problems solved by technology

But at the same time, there are still many insurmountable shortcomings in the prokaryotic expression system: for example, the commonly used expression system cannot regulate the expression time and expression level, the continuous expression of some genes may have toxic effects on the host cells, and overexpression may lead to abnormal Physiological reactions, the target protein is often expressed in the form of inclusion bodies, resulting in difficulties in product purification; and the post-translational processing and modification system of the prokaryotic expression system is imperfect, and the biological activity of the expressed product is low

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