Multi-gene binary expression vector constructed by using homologous recombination and preparation method and application of multi-gene binary expression vector

The invention relates to the field of gene engineering, in particular to a multi-gene binary expression vector constructed by using homologous recombination. The multi-gene binary expression vector is formed by connecting single genes or multiple genes into a multi-gene polymer through polyclonal enzyme cutting sites so as to form a homologous recombination intermediate vector and adding the single-gene or multi-gene polymer into a basic binary vector by homologous recombination, wherein the homologous recombination can be performed for multiple times, so that the T-DNA length of the basic binary vector is gradually increased. The invention also discloses a construction method of the multi-gene binary expression vector and application in plant transgenes. The multi-gene binary expression vector applied in plant genetic modification can transfer multiple target genes at the same time, and has the advantage of high co-transformation efficiency.