Construction method for simultaneously expressing two foreign protein vectors TRVe2 and application

Abstract

The invention discloses a construction method for simultaneously expressing two foreign protein vectors TRVe2. The construction method comprises the following steps: cloning pYL156 by agrobacterium infection of genome RNA2 of TRV as a material, constructing a vector containing 2b gene promoter and pea early-browning virus coat protein gene promoter sequences of TRV, and driving expressions of foreign genes in nicotiana benthamiana by virtue of genome promoters of the two plant viruses. The plant virus TRV2e2 capable of simultaneously expressing two non-fusion-proteins is first reported internationally; the virus does not cause any obvious symptoms in the nicotiana benthamiana, carries two foreign genes to systematically expand to the whole plant and is capable of expressing two foreign proteins in the same cell.

Description

technical field [0001] The invention relates to the field of molecular biology, and specifically includes a method for constructing and using a plant virus expression vector for simultaneously expressing two non-fused foreign proteins in the whole plant. Background technique [0002] Plant virus genomes are generally small, easy to perform molecular biology operations, and the process of virus infecting hosts is simple. Therefore, the use of plant virus vectors to express foreign genes has potential application advantages in the field of biotechnology. Plant viruses are increasingly becoming an alternative for recombinant protein expression systems, and the future development direction of plant virus expression vectors will be to use cereals, beans and other grain and oil economic crops as bioreactors to produce edible vaccines. Compared with plant transgenic production of foreign proteins, the plant virus expression system has the following advantages: first, the virus mult...

AI Technical Summary

Problems solved by technology

[0009] At present, the expression vectors of multiple proteins are constructed by using the hydrolase properties of plant virus-encoded polyproteins or the self-cleavage of FMDV peptidase 2A. The fundamental principle is to construct multiple foreign protein genes and viral genes in the same ORF After the reconstituted virus infects plants, it produces a large protein, and under the action of viral hydrolase or FMDV peptidase 2A, it produces the target foreign protein, so the foreign protein produced in plants contains a segment derived from Amino acid residues of non-target proteins, while the N-terminal or C-terminal of the target protein carries amino acid residues of other proteins that may interfere with its biological functions; viral vectors that simultaneously express two or more non-fused foreign proteins in host plants Not yet reported

Images