【摘要】 虾青素是一种具有极强抗氧化活性的类胡萝卜素,具有广泛的应用价值。β-胡萝卜素酮化酶(BKT)和β-胡萝卜素羟化酶(CrtR-B)是虾青素的生物合成途径中的两个关键酶。采用LaTaqDNA聚合酶用PCR的方法分别从pET-28a(+)-bkt,pET-28a(+)-CrtR中扩增得到bkt和CrtR-B基因,用bkt基因替换pBI221中的GUS基因形成含有CaMV 35S启动子和NOS终止子的bkt基因表达盒,然后插入植物表达载体pCAMBIA1301的多克隆位点,再用CrtR-B基因替换pCAMBIA1301中的另一个GUS基因,形成CrtR-B基因表达盒,最终获得带有抗性基因的双价植物表达载体pCAMBIA1301-bkt-CrtR。通过农杆菌EHA105介导将其转入花生(花育23),获得了经卡那霉素筛选的抗性植株,通过PCR扩增验证,花生基因组中含有这两个基因。更多还原

【Abstract】 Astaxanthin with high antioxidant activity is of great application value.BKT and CrtR-B are two key biosynthesis enzymes of astaxanthin,encoded by bkt gene and CrtR-B gene.Bkt gene expression cassette was constructed by replacing GUS gene in vector pBI221 with bkt gene by PCR method,which contained the CaMV 35S promoter and NOS terminator.Plant expression vector pCAMBIA1301-bkt-CrtR with binary gene was constructed by inserting bkt gene expression cassette into multiple cloning site and replacing GUS gene with CrtR-B gene in vector pCAMBIA1301.Bkt gene and CrtR-B gene were transferred into peanut mediated by Agrobacterium tumefaciens and under the pressure of Kanamycin selection,the kanamycin resistant transformants were achieved.The PCR amplification confirmed that bkt gene and CrtR-B gene had been integrated into the peanut genome successfully.更多还原