2A肽介导的甜菜色素多基因表达载体构建及原核表达分析  

Construction and Prokaryotic Expression Analyses of a 2A Peptide-linked Co-expression Vector for Betalains

作　　者：付瑛格 李霆格 潘庆龙 谷佳 史佑海[1] 周扬[1,2] 赵莹 王健[1] Fu Yingge;Li Tingge;Pan Qinglong;Gu Jia;Shi Youhai;Zhou Yang;Zhao Ying;Wang Jian(Key Laboratory of Genetics and Germplasm Innovation of Tropical Forest Trees and Ornamental Plants(Ministry of Education),Key Laboratory of Germplasm Resources Biology of Tropical Special Ornamental Plants of Hainan,College of Forestry,Hainan University,Haikou,570228;College of Horticulure,Hainan University,Haikou,570228)

机构地区：[1]海南大学林学院热带特色林木花卉遗传与种质创新教育部重点实验室海南省热带特色花木资源生物学重点实验室,海口570228 [2]海南大学园艺学院,海口570228

出　　处：《分子植物育种》2022年第4期1184-1190,共7页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31760590);海南省重点研发项目(ZDYF2019041);海南省自然科学基金创新团队项目(2018CXTD331);上海市资源植物功能基因组学重点实验室开放课题(PFGR202101)共同资助。

摘　　要：目前,自然界中未发现同时含有花青素和甜菜色素的植物,如能将甜菜色素导入到含花青素的植物中,则可能培育出全新植物品种。本研究以红甜菜(Beta vulgaris L.var.cicla L.)为植物材料,克隆甜菜色素合成途径中BvDODA1和BvCYP76AD1两个关键基因,利用重叠PCR(splicing by overlap extension PCR,SOEPCR)技术将这两个基因通过FMDV(Foot and mouth disease virus)2A肽串联得到融合基因,命名为DAC,并在大肠杆菌中进行了表达分析验证。结果表明,BvDODA1基因可在大肠杆菌中正常表达发挥功能,而BvCYP76AD1基因在大肠杆菌中未能正常发挥功能。本研究结果为后期甜菜色素融合基因转化花青素植株、培育新品种以及利用微生物发酵生产甜菜色素提供了理论依据。Plants with both of betalains and anthocyanin have not been found in nature at present.If betalains can be introduced into anthocyanin-generated plants,some new cultivars with innovative colors can be breed.The study cloned the key genes of BvDODA1 and BvCYP76AD1 in the betalains synthesis pathway using the red beet(Beta vulgaris L.var.cicla L.)as the plant materials,and contructed the BvDODA1 and BvCYP76AD1 fusion gene linked by FMDV 2A peptide(named as DAC gene)with splicing overlap extension PCR method(SOE-PCR).The expression analysis of DAC was performed in Escherichia coli.The results showed the BvDODA1 gene in pETDAC vector could express in E.coli.However,the function of BvCYP76AD1 was failed in E.coli.The results of this study provide theoretical basis for the transformation of betalains into anthocyanin-generated plants,the breeding of the new varieties and the production of betalains by microbial fermentation.

关 键 词：甜菜色素 2A肽 SOE-PCR 原核表达 

分 类 号：S566.3[农业科学—作物学]