白屈菜CYP719A13基因的克隆及植物转化载体构建  

Cloning of CYP719A13 Gene from Chelidonium majus L.and Construction of Plant Transformation Vector

作　　者：洪波[1,2] 朱雅静 许永华[1] 赵露[1] Hong Bo;Zhu Yajing;Xu Yonghua;Zhao Lu(College of Traditional Chinese Medicine,Jilin Agricultural University,Changchun,130118;College of Resource and Environmental Science,Jilin Agricultural University,Changchun,130118)

机构地区：[1]吉林农业大学中药材学院,长春130118 [2]吉林农业大学资源与环境学院,长春130118

出　　处：《分子植物育种》2024年第23期7750-7756,共7页Molecular Plant Breeding

基　　金：吉林省教育厅“十三五”科学技术项目(JJKH20200365KJ);吉林省科技厅科技发展计划项目(20210203130SF)共同资助

摘　　要：基于本课题组前期的研究发现,白屈菜CYP719A13基因表达量与血根碱、白屈菜红碱等生物碱含量呈现显著正相关,是合成路径中一种非常关键的酶基因。本研究选用pMD-19-T作为克隆载体,并转化导入大肠杆菌DH5α,对白屈菜CYP719A13基因进行了克隆,通过基因测序方法验证了该基因。构建了植物表达载体pRI201-CYP719A13,并通过电击法转化导入农杆菌LBA4404,利用菌液PCR法验证了该表达载体构建成功。本研究为后续研究该基因功能,在利用分子育种手段提高白屈菜中生物碱含量方面发挥作用提供了基础。Based on the previous research of our group,it was found that CYP719A13 gene expression had a significant positive correlation with the content of alkaloids such as sanguinarine,chelerythrine and other alkaloids in Chelidonium majus.It was a very key enzyme gene in the synthesis path.In this study,PMD-19-T was used as the cloning vector and transformed into escherichia coli DH5αto clone the CYP719A13 gene,which was verified by gene sequencing.The plant expression vector pRI201-CYP719A13 was constructed and transformed into Agrobacterium LBA4404 by electroshock method.The successful construction of the expression vector was verified by bacterial solution PCR method.This study provided a basis for further research on the function of this gene,which may play an important role in the increasing the alkaloid content of Chelidonium majus by molecular breeding.

关 键 词：白屈菜 CYP719A13基因 表达载体构建 

分 类 号：S567.239[农业科学—中草药栽培]