Plant expression plasmid vector containing C-Myc protein fusion label and construction method for vector

Abstract

The invention discloses a plant expression plasmid vector containing a C-Myc protein fusion label. The plant expression plasmid vector contains the C-Myc protein fusion label at a terminal C of the plant expression plasmid vector. The construction method for the plasmid vector comprises the following steps: (1) taking pCAMBIA1302 as a departing plasmid, carrying out restriction enzyme digestion onthe plasmid by using SpeI and Eco065I, and recycling a klenow fragment through electrophoretic gel cutting; (2) carrying out full-sequence synthesis on a nucleotide sequence encoded with three C-Mycproteins; (3) designing an amplification primer; and (4) adding a SpeI restriction enzyme cutting site at a terminal N, and adding an Eco065I restriction enzyme cutting site at the terminal C; carrying out restriction enzyme digestion on the synthesized fragment by using SpeI and Eco065I; and finally connecting products, transforming DH5 alpha by heat shock, and carrying out PCR identification toobtain positive clone. The plant expression plasmid vector has the advantages that the plant expression plasmid vector of a fusion label of small protein is obtained, a user is assisted to rapidly research functions of the protein, functions of the target protein cannot be interfered, and it ensures that the functions of the protein can be researched accurately.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a plant expression plasmid vector containing a C-Myc protein fusion tag and a method for constructing the vector. Background technique [0002] C-Myc tag protein is a small tag containing 10 amino acids, the tag sequence Glu-Gln-Lys-Leu-Ile-Ser-Glu-Glu-Asp-Leu, these 10 amino acids are expressed as antigenic epitopes in different Its corresponding antibody is still recognized in the protein frame. C-Myc fusion protein tag has been successfully applied in Western-blot hybridization, immunoprecipitation, and chromatin immunoprecipitation to analyze protein expression, protein interaction, and protein regulation of target genes. However, small protein fusion tags such as C-Myc will not or rarely interfere with the function of the fused protein. In order to scale and industrialize the C-Myc fusion protein tag, it is necessary to provide a plant expression plasmid vector w...

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Problems solved by technology

[0003] In order to solve the problem that the pCAMBIA carrier does not fuse with the small protein fusion tag of C-Myc in the prior art, the invention provides a plant expression plasmid vector containing the C-Myc protein fusion tag and a method for constructing the carrier thereof, to realize The purpose is to obtain a plant expression plasmid vector containing a fusion tag of a small protein such as C-Myc protein, so as to help users quickly study protein functions without interfering with the function of the target protein itself, ensuring that it can Accurately study protein function

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