拟南芥CBF1基因植物表达载体构建及其对香蕉遗传转化.pdfVIP

451 (1. 510640 2. 410128 ) * PCR CBF1(CRT/DRE binding factor 1)CBF1 (NM_118681) 100%CBF1 pBI121 Xba I Sca I pBI121-CBF1 EcoR I Hind III 35S CBF1 pBI121-CBF1 pCAMBIA1301 p1301-CBF1 EHA105 (Musa ABB cv. dajiao)GUS CBF1 CBF1 Construction of Plant Expression Vector of CBF1 and its Genetic Transformation of Banana 1,2 1* 1 1 Liu Kai , Hu Chunhua , Wei Yuerong , Yi Ganjun 1 2. ( .Fruit Research Institute, Guangdong Academy of Agricultural Science, Guangzhou, 510640; Horticulture and Landscape CollegeHunan Agriculture University, Changsha,410128) *This author contributed equally to this work Abstract:The CBF1 gene(CRT/DRE binding factor 1) was cloned from Arabidopsis thaliana by using PCR, The result of sequence analysis showed that the cloned gene has 100% similarity with that of the CBF1 gene from Arabidopsis thaliana(accession number: NM_118681) deposited in Genbank. Then, the CBF1 gene were excised from pMD18-T cloning vector with restriction enzyme Xba and Sca I and inserted into the site between the same two restriction enzymes on pBI121 vectors to abtain the middle recombinant plasmid pBI121-CBF1. After that, The fragments containing 35S-CBF1 were cut from recombinant plasmid pBI121-CBF1 by EcoR I and Hind III, and directionally linked it to the plant expression vector pCAMBIA1301 to get the plant expression vector p1301-CBF1. we did transform of the embryogenic cell suspensions of Dajiao(Musa ABB cv. dajiao) via Agrobacterium -mediated transformation. The