博落回抗生素敏感度筛选与植物表达载体构建  

Screening of Antibiotic Sensitivity and Construction of Plant Expression Vector in Macleaya cordata

作　　者：徐敏 夏丽琼[2,3] 黄鹏[1,2] 唐其 曾建国[1,2,3] 刘薇[2] 

机构地区：[1]湖南农业大学园艺园林学院,长沙410128 [2]湖南农业大学兽用中药资源与中兽药创制国家地方联合工程研究中心,长沙410128 [3]湖南中医药大学药学院,长沙410128

出　　处：《分子植物育种》2018年第4期1133-1137,共5页Molecular Plant Breeding

基　　金：湖南省科技重点计划(2016SK3002);湖南省教育厅科学研究项目(16C0769);湖南省自然科学基金项目(2016JJ4040)共同资助

摘　　要：采用不同浓度抗生素对博落回组培苗叶片外植体处理,并对比不同浓度的卡那霉素、潮霉素、头孢和特美汀的筛选效果。结果表明,卡那霉素比潮霉素更适合用于博落回外植体的筛选,其最适宜浓度为40 mg/m L。特美汀比头孢更适合用于农杆菌的抑制,其最适宜浓度为400 mg/m L。除此以外,还利用In-Fusion技术将线性载体与博落回BBE基因片段连接,测序结果与预期序列一致,表明成功构建了p CAMBIAD-BBE植物表达载体。该工作为优化博落回遗传转化体系和构建过表达博落回植株提供研究依据。The study used different concentrations of antibiotics to treat leaf explants of Macleaya cordata, and compared the screening effects ofkanamycin, hygromycin, Cefotaxime and Timentin with different concentrations. The results showed that kanamycin was more suitable for the screening of Macleaya cordata explants than hygromycin, and its optimum concentration was 40 mg/mL. Timentin is more suitable for the inhibition of Agrobacterium than Cefotaxime, and the most suitable concentration was 400 mg/mL. In addition, the linear vector was ligated with the BBE gene fragment in Macleaya cordata by In-Fusion technique, and the sequencing results were consistent with the expected sequence, indicating that the pCAMBIAD-BBE plant expression vector was successfully constructed. This work laid the foundation for the optimization of the genetic transformation system and the construction of over-expressive Macleaya cordata plant.

关 键 词：博落回 抗生素 载体构建 

分 类 号：Q943.2[生物学—植物学]