关键词: WIN1基因, 基因克隆, 烟草, 农杆菌

Abstract: The WIN1 genes are related to the process of gene expression in waxy metabolism. In this research, the total RNA was extracted from the wild type arabidopsis buds and amplified into SHN1 WIN1. The plant expression vector PBI121-SHN1/WIN1 was constructed and the WIN1 genes were inserted into tobacco k326 by using the agrobacterium-mediated technique. There were four different culture media in this research, the minimal medium was T1: MS+1 mg·L^-1 6-BA+0.1 mg·L^-1 NAA; the medium for filtration sterilization was T2: MS+1 mg·L^-1 6-BA+0.1 mg·L^-1 NAA+500 mg·L^-1 Carb; the medium for screening tobacco was T3: MS+100 mg·L^-1 Kan; the medium for tobacco subculture was T4: MS+80 mg·L^-1 Kan+300 mg·L^-1 Carb; the medium for rooting culture of tobacco was T5: MS+0.1 mg·L^-1 Naa+300 mg·L^-1 Carb+80 mg·L^-1 Kan. In this experiment, the WIN1 genes were inserted into tobacco k326 by the plant genetic engineering to change the total gene sequences of tobacco k326. By the detection of PCR and RT-PCR, the WIN1 genes were successfully inserted into tobacco k326, and eventually the transgenic tobacco k326 were obtained. In this experiment, the WIN1 genes were inserted into tobacco k326 through the plant genetic engineering and to make the cutin membrane become thick and the composition changes are expected. Another object was to increase the drought-resistance, cold-tolerance and disease resistance of tobacco K326, therefore, the resistance of tobacco can be improved. The experiment is not only meaningful to reveal the function of plant cutin membrane structure, but also has great important application value in plant gene engineering technology and cultivate quality variety of plants and agricultural production.

Key words: WIN1 genes, gene clone, tobacco, agrobacterium