【作者】 王宏华；

【导师】 郭慧云； 秦松；

【作者基本信息】 吉林大学 ， 生物化学和分子生物学， 2005， 硕士

【摘要】 水稻蜡质基因(Waxy)编码结合在淀粉粒上的淀粉合成酶,它控制花粉、胚乳与胚囊中直链淀粉的合成,是一种组织与发育阶段专一性表达的基因。本研究工作通过基因枪转化技术连接报道基因GUS 的水稻蜡质基因启动子载体导入玉米愈伤组织,取转化后48h 玉米幼胚,GUS 染色后作石蜡切片,肉眼即可见大面积蓝色斑块,转化后45d 玉米愈伤组织GUS 染色后作石蜡切片仍然可看到蓝色斑块,未转化玉米均未见蓝色斑块。显示水稻Wx 基因启动子可以驱动GUS 基因在发育的玉米幼胚和愈伤组织中表达。CaMV35S 启动子是植物中常用到的组织非特异性启动子,水稻蜡质基因启动子是胚乳特异性启动子。将β-胡萝卜素酮化酶基因(bkt)分别连入这两个启动子下,引入玉米后,一方面可以比较启动子不同造成转基因玉米类胡萝卜素的各个组分的差异,另一方面也可论证玉米种子在水稻蜡质基因启动子作用下产虾青素的能力,开发其多种用途。更多还原

【Abstract】 Tissue-specific promoter can drive foreign genes’expression in plant’s specific tissue and specific time.Rice wary gene encodes the starch synthesizing enzyme combining to starch,it controls the synthesizing of amylase in the farina、endosperm and megaspore.The rice wary gene promoter and GUS gene were linked and transformed to maize type II callus. Many blue spots were observed in the transgenic maize callus by histochemical staining, however there was no in the comparision.45 days later ,the blue sports can still be seen by histochemical staining.The results suggest that rice wary gene promoter an drive foreign genes’expression in maize endosperm. Furthermore, foreign genes’expression in the different part of maize will be checked to see if it is exist and if it is strong or unconspicuous by steady heredity. Early nptII gene was used for screening of the transgenic maize.Now it is found that there is natural resistive in the maize tissue to the nptII gene.So nptII gene is gradually substituted by hpt and pat gene. It is proved that maize calli was sensitive to Hygromycin, 5μg/ml Hygromycin will obviously restrain the growth of maize callus.With the increasing of Hygromycin concentration,the death of maize calli increases.When the Hygromycin concentration is 50μg/ml, all the maize calli will die.The half death is 20μg/ml. There is much Zeaxanthin in the maize but not find Astaxanthin.It is known from the synthesizing way of isoprenoids that Zeaxanthin being catalyzed by bkt gene may be made into Astaxanthin. This research bkt gene was linked with CaMV35S promoter or rice waxy gene promoter更多还原