Plasmid system for constructing plant multi-gene expression vector and application of plasmid system

Abstract

The invention discloses a plasmid system for constructing a plant multi-gene expression vector and an application of the plasmid system. By transforming an escherichia coli plasmid vector pUC18 and a dual-core agrobacterium Ti plasmid pBI121, recombinant plasmids pKAFCR0 and pKAFCR100 are formed. The pKAFCR0 has a 35S promoter and an NOS terminator which are the most common in plant gene expression, a plurality of cloning restriction sites are introduced to the upstream side, the middle side and the downstream side of the pKAFCR0, and a gene segment, which is obtained by conducting cloning by virtue of such methods as PCR and the like, can be linked to the middle part of the 35S and the NOS of the pKAFCR0 by virtue of a plurality of modes; and the pKAFCR100 has a kanamycin NPT II tolerance gene and an sGFP green fluorescent protein reporter gene, as well as multiple-cloning restriction sites in middle. By using the pKAFCR0 and pKAFCR100 plasmids in a matched mode, one or more plant gene expression vector can be conveniently constructed.

Description

technical field [0001] The invention relates to the field of biology, in particular to a plasmid system for constructing a plant multigene expression vector and its application. Background technique [0002] The rapid development and wide application of plant genetic engineering technology has opened up broad prospects for the genetic improvement of plants. This technology overcomes the limitations of plant sexual hybridization, and can introduce genes from different species or even artificial synthesis into plants, thereby improving plant traits and cultivating new varieties of high-quality and high-yield crops. In the research and application of molecular biology such as gene cloning, gene function analysis, and gene transformation, vector construction is a commonly used technical means, and the use of appropriate expression vectors and construction methods will make the experimental effect twice the result with half the effort. However, plasmids currently commonly used i...

AI Technical Summary

Problems solved by technology

Although TOPO cloning is simple, the range of target vectors is limited. Gateway and infusion PCR can ignore restriction sites, but the technologies and vectors developed by these companies are mainly aimed at the construction of gene expression vectors for animals and microorganisms, and lack plant gene expression. The necessary functional elements such as promoters, terminators, and screening markers are relatively expensive (about 3,000-10,000 yuan), and due to patent restrictions, they can only be used for research purposes, which is limited in application

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