Easy and efficient agrobacterium tumefaciens induced corn gene transfer method
An Agrobacterium-mediated, transgenic technology, applied in plant genetic improvement, botanical equipment and methods, applications, etc., can solve the problems of tissue culture, callus differentiation difficulties, application limitations, and low transformation efficiency, and achieve unique application value. , Easy to operate, avoid the effect of complicated process
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[0010] The technological process of the present embodiment method is as follows:
[0011] Exogenous gene→Ti plasmid expression vector construction→Agrobacterium transformation solution preparation→in situ transformation→transformant screening→transformant identification→transgenic plants.
[0012] The specific implementation steps are as follows:
[0013] 1. Determination of the target gene and vector construction.
[0014] The relevant target gene is selected, and the maize expression vector is constructed using Ti plasmids such as pCAMBIA1300 or pBI121.
[0015] 2. Preparation of Agrobacterium transformation solution.
[0016] The expression vector is introduced into the competent cells of Agrobacterium by a three-parent hybridization method or a direct transformation method to obtain transformed Agrobacterium strains.
[0017] (1) Three-parent hybridization method, each of the three cultured bacterial solutions of Agrobacterium tumefaciens LBA4404, Escherichia coli HB101...
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