虾青素合成关键酶基因bkt植物表达载体的构建 及对玉米的遗传转化

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）是由玉米黄素合成虾青素生物合成途径中的关键酶。采用La Taq DNA聚合酶用PCR的方法从pET-28a（+）bkt中扩增得到bkt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，最终获得带有选择标记和报告基因的植物表达载体pCAMBIA1301-bkt。通过农杆菌LBA4404介导将其转化进入玉米自交系齐319，转化后的愈伤经GUS组织化学染色分析表明bkt基因已经转入玉米胚性愈伤组织

Construction of Plant Expression Vector Containing Genes Encoding The Key Biosynthesis Enzyme of Astaxanthin( bkt) and Its Expression in Zea mays L.

Astaxanthin with high antioxidant activity is of great application value. Bkt is the key biosynthesis enzyme of astaxanthin which encoded by bkt gene. Bkt gene expression cassette was constructed by replacing GUS gene in vector pBI221 with bkt gene obtained by PCR method, which contained the CaMV 35S promoter and NOS terminator. Plant expression vector pCAMBIA1301-bkt was constructed by inserting bkt gene expression cassette into multiple cloning site in vector pCAMBIA1301.Bkt gene was transferred into maize Qi 319 mediated by Agrobacterium tumefaciens. The results of GUS histochemical assay indicated that bkt gene had been transferred into maize emberyogenic calli.