摘要
来源于萤火虫的荧光素酶结构基因与花椰菜花叶病毒35SRNA启动子以及胭脂碱合成酶DNA3’末端结构相拼接,构成一融合基因.通过叶盘感染法将该融合基因引入烟草叶盘,把选择诱导培养基上形成的愈伤组织转到选择分化培养基后能很快分化出芽点,继而长成完整的再生植株.对再生植株提取物发光试验表明,外源基因已整合到烟草细胞的核基因组上,并能在烟草中表达.带有这种融合基因的重组质粒pBC341具有KpnI、SstI等单一的酶切位点,因而也可作为常规的基因载体.
Firefly luciferase gene encodes an enzyme that catalyzes the adenosine triphos-phate-dependent oxidation of luciferin and thus produces luminescence. The authors constructed a chimeric luciferase gene which was promoted by the CaMV (cauliflower mosaic virus) 35 s RNA promoter and spliced with 3'terminal structure of carmine synthetase DNA. The chimeric luciferase gene was introduced into the leafplate of tobacco by leafplate infection method. The calli originated from selective induction medium can differentiate into buds quickly on selective differential medium and finally develop into whole plants. The luciferase activity assay indicated that the foreign gene had been integrated into the genome of tobacco and could be expressed in plant. The luciferase gene can be particularly valuable in phytogenetics as a marker for cell transformation. The recombinant plasmid pBC341 has not only the chimeric luciferase gene but also some unique restriction enzyme sites like Kpn I and Sst I , into which the foreign genes can be easily inserted so that it can be used as a conventional gene vector.
关键词
荧光素酶
基因
转化
表达
烟草
luciferase gene
tobacco
transformation
expression
