大肠杆菌otsB基因的克隆与转化本生烟草的初步研究  

Preliminary study on cloning of otsB gene in E.coli and its transformation into Nicotiana benthamiana

作　　者：陆玉建[1,2,3] 张韩杰[1] 刘南南[1] 韩文瑜[3] 沈志强[2] 

机构地区：[1]滨州学院生命科学系/山东省黄河三角洲野生植物资源开发利用工程技术研究中心,山东滨州256603 [2]山东省滨州畜牧兽医研究院博士后科研工作站,山东滨州256600 [3]吉林大学博士后科研流动站,吉林长春130062

出　　处：《广东农业科学》2015年第8期108-112,193,共5页Guangdong Agricultural Sciences

基　　金：山东省自然科学基金(ZR2012CL14);滨州学院博士基金(2010Y08)

摘　　要：海藻糖是一种非还原糖类,可作为渗透调节剂,提高生物在非生物胁迫条件下的抗逆能力。6-磷酸海藻糖磷酸化酶(TPP)是海藻糖生物合成中的重要酶类,在大肠杆菌中,ots B基因编码TPP。以大肠杆菌的基因组DNA为模板,通过菌落PCR技术克隆得到ots B基因。将目的基因和p2300-GFP相连,从而构建p2300-ots BGFP表达载体。利用根癌农杆菌介导的叶盘法转化本生烟草,将ots B导入到受体细胞中。通过组织培养和抗生素筛选,初步获得具有抗性的转基因本生烟草。Trehalose is a non-reducing sugar, which can be used as osmotic regulator to improve the resistance of plants under abiotic stress conditions. TPP is the key enzyme for trehalose biosynthesis and otsB gene encodes TPP in E.coli. In this experiment, the genomic DNA of E.coli was as template to clone otsB gene by PCR method. Then the target gene was connected with p2300-GFP plasmid, which would lead to the construction of p2300-otsB-GFP vector. The p2300-otsB-GFP expression vector was transformed into Nicotiana benthamiana using leaf disc method mediated by Agrobacterium tumefaciens. After that the transgenic N. benthamiana was preliminarily obtained through the tissue culture and antibiotics screening.

关 键 词：海藻糖 otsB基因 表达载体 本生烟草 根癌农杆菌 

分 类 号：S572[农业科学—烟草工业]