白屈菜CmFAD2基因的克隆及植物转化载体构建  

Cloning of CmFAD2 Gene from Chelidonium majus L.and Construction of Plant Transformation Vector

作　　者：赵露[1] 李俊 周新芳 苑雪琪 朱雅静 魏云洁[2] 许永华[1] Zhao Lu;Li Jun;Zhou Xinfang;Yuan Xueqi;Zhu Yajing;Wei Yunjie;Xu Yonghua(College of Traditional Chinese Medicine,Jilin Agricultural University,Changchun,130118;Institute of Special Products,Chinese Academy of Agricultural Sciences,Changchun,130000)

机构地区：[1]吉林农业大学中药材学院,长春130118 [2]中国农业科学院特产研究所,长春130000

出　　处：《分子植物育种》2020年第6期1876-1882,共7页Molecular Plant Breeding

基　　金：吉林省科技发展计划项目(20190304023YY)资助。

摘　　要：通过对白屈菜低温应答过程的转录组分析发现膜脂不饱和化相关基因的表达在一定过程中发生变化,脂肪酸去饱和酶基因FAD2在随温度的变化趋势为正"V"型,且表达量变化显著。利用NCBI等在线软件对序列进行相关生物学信息分析,并对白屈菜FAD家族成员FAD2基因的完整开放阅读框(ORF)进行克隆,并命名为CmFAD2。选用克隆载体pMD-19-T,转化大肠杆菌DH5α,测序验证序列正确性及完整性。将目的基因与植物表达载体pRI-201-AN连接构建重组DNA pRI-201-AN-Cm FAD2,电击法转化农杆菌LBA4404,利用菌液PCR法验证成功。该基因可作为药用植物抗寒品种创制的候选基因。The transcriptome analysis of low temperature response in Chelidonium majus L.showed that the expression of genes related to membrane lipid unsaturation changed in a certain process.Fatty acid desaturase gene FAD2 has a positive"V"type with a significant change in expression with temperature.The sequence was subjected to relevant biological information analysis using online software such as NCBI,and the complete open reading frame(ORF)of the FAD2 gene,a member of the FAD family of celandine was cloned and named as CmFAD2.The cloning vector pMD-19-T was used and transformed into E.coli DH5α.The sequence was verified for correctness and integrity.The target gene was ligated with the plant expression vector pRI-AN-201 to construct recombinant DNA pRI-201-AN-CmFAD2.It was transformed into Agrobacterium LBA4404 by electroporation and verified by bacterial solution PCR.The gene can be used to judge cold-resistant varieties of medicinal plants.

关 键 词：白屈菜 低温 CmFAD2 表达载体构建 

分 类 号：S567.239[农业科学—中草药栽培]