GNA基因表达载体的构建及遗传转化甘蔗  

Construction of Plant Expression Vector of gna Gene and Transformation in Sugarcane

作　　者：刘晓娜[1,2] 冯翠莲[1,2] 张树珍[1] 

机构地区：[1]中国热带农业科学院热带生物技术研究所,海南海口571101 [2]海南大学农学院,海南儋州571737

出　　处：《热带作物学报》2010年第6期887-893,共7页Chinese Journal of Tropical Crops

基　　金：中央级公益性科研院所基本科研业务费(No.ITBBZD0724);中国热带农业科学院院基金(No.Rky0617);现代农业产业技术体系建设专项基金(No.nycytx-24)资助

摘　　要：利用经人工改造的gna基因分别与Ubi、Rbcs启动子组合,构建抗虫植物表达载体pUNG和pRNG,通过农杆菌介导法导入甘蔗愈伤组织,经PPT筛选和PCR检测,获得Ubi-gna转基因植株124株,Rbcs-gna转基因植株35株。对部分转基因植株进行RT-PCR检测,初步证明外源基因已经整合到甘蔗基因组中,并得到有效表达;对RT-PCR阳性植株进行GNA(植物外源凝集素)活性测验,结果表明,甘蔗叶片表达的凝集素可以使健康公鸡的红细胞凝集,初步证明表达的GNA具有正常的生物学活性。The plant expression vectors pUNG and pRNG,harboring gna gene driven by Ubi and Rbcs promoter respectively was constructed and introduced into sugarcane by Agrobaterium-mediated transtormation,and 124 Ubi-gna transgenic plants and 35 Rbcs-gna transgenic plants were obtained after successive PPT resistance selection and PCR detection.Some of the transgenic plants had been detected by RT-PCR analysis,proved that the foreign gene had been integrated into sugarcane genome and expressed.The agglutinant ex tracted from the leaves of the positive transgenic plants could agglutinate the red blood cells from healthy cock,proving that the expressed gna gene had normal biological activity.

关 键 词：雪花莲凝集素 GNA基因 PCR 

分 类 号：S566.1[农业科学—作物学] Q78[生物学—分子生物学]