RNAi Vector Construction Of AtDof1.7 Gene And The Transformation Of Arabidopsis Thaliana

Gene expression is regulated in living cells through different mechanisms, although the most important one is transcriptional control.Transcription factors play important roles in regulation of plant growth and development, as well as its response to environment, through the activation or inhibition effects on the target genes The Dof (DNA-binding with one finger) transcription factors are the member of a family of plant-specific transcription factors that have a highly conserved DNA-binding domain and play an important role in regulating the growth and development of the plants.In order to study the regulation of AtDofl.7 gene in fatty acid metabolic pathway of Arabidopsis thaliana seeds, AtDofl.7 gene cloned was inserted to the both sides of Chalcone sythase intron of pFGC5941 plasmid, with a seed specific expression NapinA promoter instead of CaMV35S, in forward and reverse direction, respectively, to construct a AtDofl.7 gene RNAi vector which was used to interfere the gene expression.Then the RNAi vector pAdofl was transfered into Arabidopsis thaliana by Agrobacterium-mediated. Transformants were screened by PPT resistance and PCR amplification. Finally,5 positive transformation of seedlings were detected. Utilizing Gas chromatography,the T2 generation seeds were analyzed which showed fatty acid ratio of seeds were changed, comparison with wild-type Arabidopsis thaliana seeds.The Oleic acid increased,but Linolenic acid decreased in the transgenic seeds,which suggested AtDofl.7 gene maybe regulates the fatty acid metabolic pathway of Arabidopsis thaliana seeds.