Method for constructing a universal plant expression vector

Abstract

The invention provides a method for constructing a universal plant expression vector, and belongs to the technical field of organism. The method comprises the steps of constructing an intermediate vector containing a promoter terminator; connecting a target gene to an efficient plant transformation vector pCMBIA1300 by the intermediate vector to obtain the plant expression vector; transforming the constructed plant transformation vector into a rice cell, screening the transformed regeneration plant (T0 generation) from explants in a plant regeneration medium, and detecting a transformation system to obtain the transgenic rice. The method disclosed by the invention can construct the plant expression vector by double polyclonal restriction points of the vector, so as to expand the application range of the method.

Description

technical field [0001] The invention relates to a method for constructing a universal plant expression vector, which belongs to the field of biotechnology. Background technique [0002] In plant genetic engineering research, constructing plant expression vectors is a very important link. A vector is a medium that carries target DNA fragments into host cells for amplification and expression. Without a vector, the target gene cannot enter the recipient, and even if it enters the recipient cell, it cannot be expressed. Bacterial plasmid vectors or phage vectors are connected with promoter genes and gene sequences of polyribosomes to form expression vectors. The construction of expression vectors is a very important but cumbersome work, therefore, the research on the construction methods of expression vectors has important practical significance. [0003] With the development of plant genetic engineering technology, various vector systems suitable for different research purpos...

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Problems solved by technology

[0008] The above are several methods for constructing plant expression vectors. Among them, the traditional construction method is universal and unlimited, but the construction efficiency is not stable. It is necessary to select a suitable site in the process of enzyme digestion and ligation. Once there is no suitable site, a simple The construction of the

Gateway technology is also relatively general, and vector construction can be completed through two reactions, which is extremely efficient, but it often needs to be combined with other methods to obtain entry clones, and its cost is high

The three-segment T-DNA method requires Agrobacterium-mediated co-transformation method to remove the selection marker, and can obtain safe transgenic plants, but it generally requires field screening and isolation, which is only suitable for the purpose vector of transgenic research, and needs to obtain seeds for the next generation Plant separation to obtain unmarked plants

The complex vector construction method is suitable for the construction of large fragment vectors, with relatively few steps and high efficiency, but has no advantages for the construction of some simple vectors. At the same time, it needs to be combined with PCR, which also limits its application range

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