Cloning Of BnGRF2 And Glyphosate Resistance EPSPs Gene And Construction Of The Plant Bivalent Expression Vector

China is the world’s largest producer of rape. Although the development of rapeseed industry has made a great achievement in our country, it still face many challenges and the international competition is getting fiercer. So the main research areas of rape breeding focused on increasing yield and oil content of rapeseed and curbing labour costs. GRF(Growth-regulating factor) is the transcriptional regulator and is involved in regulating the growth and development in plants. Some research has proved that Bn GRF2 could increase oil content and seed weight in transgenic Arabidopsis. Harmfulness weeds is one of the limiting factors in the development of rapeseed production, and the rural labor shortage has become more severe in china.Thanks to herbicide resistant GM crops, it becomes the more convenient way to solve the paradox. Therefore, the Bn GRF2 and herbicide-resistant gene(EPSPs)are introduced to the oil crops simultaneously. As a result, it not only reduces the production cost, but also increase the oil content and crop yield. Furthermore, it make us understanding the function of Bn GRF2 in seed development and oil metabolism, effect of bivalent genes(EPSPs, Bn GRF2) on growth of brassica napus. Also, it has some practical significance for rapeseed germplasm innovation of herbicide resistance and high oil content. The main contents of research and results are as follows.1. Cloning of seed-specific promoters Napin and Bn GRF2 gene. According to the sequence of Napin and Bn GRF2 in Genebank, the primers were designed to clone Napin and Bn GRF2 gene. Similarity comparisons show that they share 99.03% and 99.0% similarity in nucleotide sequence, it showed that the two genes were cloned successfully. The Plant CARE and PLACE were used to anlyse the Napin sequence. It showed that the Napin promoter had endosperm-specific expression elements GCN4 motif and Skn-1 motif,it also had ABA-induced and regulating element ABRE.2. Construction of plant expression vector. Restriction enzyme digestion and DNA ligation was used in the whole precess. First, Seed-specific promoter Napin gene was ligate into vector which contains bivalent genes(EPSPs), we obtained a recombinant plasmid p CENP. Then Nos terminator was inserted into vector p CENP,we got the recombinant plasmid p CENN. Last but not the least, Bn GRF2 gene was inserted into vector p CENN, the seed-specific expression vector p CEGRF was constructed which contained EPSPs gene and Bn GRF2 gene. This bivalent vector can applied to genetic improvement of seed oil content of energy plants.