盐藻GPDH基因的载体构建及在烟草中的表达

Vector Construction and Expression in Tobacco of GPDH Gene from Dunaliella salina

作者用冻融法,将构建有盐藻3 磷酸甘油脱氢酶基因的植物高效表达载体DsGPDH导入感受态的根癌农杆菌EHA105中.叶盘法转化烟草,在含卡那霉素的MS培养基上进行选择性培养,生根率达到80%.用PCR的方法对再生苗进行了初步筛选,阳性率约为50%.对PCR阳性苗,进行RT PCR分析,证实整合到烟草基因组中的GPDH基因表达产生了mRNA.

The plant transformation vector DsGPDH,which was constructed to express the glycerol-3-phosphate dehydrogenase gene was introduced into competent Agrobacterium tumefaciens EHA105 using the freeze-thaw method.After infected by A.tumefaciens,the leaf discs of tobacco were selected on plates containing MS inducement medium supplemented with 100mg/L kanamycin,the rate of rootage was 80%.The regenerated plants were identified.About 50% of tested plants produced the target band by PCR.RT-PCR analysis showed that GPDH gene integrated into tobacca genome has produced mRNA.