| 陆玉建，张韩杰，刘南南，等.大肠杆菌 otsB基因的克隆与转化本生烟草的初步研究[J].广东农业科学,2015,42(8):101-107 |
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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 andits transformation into Nicotiana benthamiana |
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| DOI： |
| 中文关键词: 海藻糖 otsB基因 表达载体 本生烟草 根癌农杆菌 |
| 英文关键词: trehalose otsB gene expression vector Nicotiana benthamiana Agrobacterium tumefaciens |
| 基金项目: |
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| 摘要点击次数: 2132 |
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| 中文摘要: |
| 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， otsB 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otsB 基因。将目的基因和p2300-GFP 相连，从而构建 p2300-otsB-
GFP 表达载体。利用根癌农杆菌介导的叶盘法转化本生烟草，将otsB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。 |
| 英文摘要: |
| Trehalose is a non-reducing sugar，which can be used as osmotic regulator to improve the resistanceof plants under abiotic stress conditions. TPP is the key enzyme for trehalose biosynthesis and otsB gene encodes TPPin E.coli. In this experiment，the genomic DNA of E.coli was as template to clone otsB gene by PCR method. Then thetarget 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 mediatedby Agrobacterium tumefaciens. After that the transgenic N. benthamiana was preliminarily obtained through the tissueculture and antibiotics screening. |
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