基于Nimble Cloning的新型植物BiFC载体的构建和应用  

Construction and Application of Novel Plant BiFC Vectors Based on Nimble Cloning

作　　者：曾妍静 沈文涛[2] 庹德财[2] 黎小瑛[2] 言普[2] 高新征 周鹏[1,2] Zeng Yanjing;Sheng Wentao;Tuo Decai;Li Xiaoying;Yan Pu;Gao Xinzheng;Zhou Peng(College of Tropical Crops,Hainan University,Haikou,570228;Hainan Academy of Tropical Agricultural Resource,Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Science,Haikou,571101;School of Basic Medical Science,Hainan Medical University,Haikou,571199)

机构地区：[1]海南大学热带作物学院,海口570228 [2]中国热带农业科学院热带生物技术研究所,海南热带农业资源研究院,海口571101 [3]海南医学院基础医学与生命科学学院,海口571199

出　　处：《分子植物育种》2019年第23期7757-7763,共7页Molecular Plant Breeding

基　　金：中国热带农业科学院基本科研业务费专项(1630052019004; 1630052016005)资助

摘　　要：Nimble Cloning是一项新型的分子克隆技术,具有操作简单、使用灵活、克隆效率高、标准化克隆等优点,但Nimble Cloning需要使用配套的表达载体。本研究将pDOE系列的4个整合型BiFC载体上的SfiI位点去除,然后在这4个载体的多克隆位点处插入NC克隆框,构建了一套适用于Nimble Cloning的新型植物BiFC表达载体,命名为pNC-BiFC-VD系列载体。将pNC-BiFC-VD载体应用到植物蛋白互作研究中,分析了番木瓜eIFiso4E和病毒PRSV的vpg蛋白之间的互作,结果表明番木瓜eIFiso4E和PRSV-vpg在细胞核发生互作。该套新型植物BiFC表达载体有望在植物蛋白互作研究中得到广泛应用。Nimble Cloning is a novel technology for molecular cloning.It is simple,versatile,and efficient.Nimble Cloning requires supporting expression vectors.In this study,the SfiI sites in the four integrated BiFC vectors of pDOE series were eliminated,and two NC Cloning frames were inserted at the multiple cloning sites of the four vectors.A new type of plant BiFC expression vectors based on Nimble Cloning were constructed and named as pNC-BiFC-VD series vectors.The pNC-BiFC-VD vectors were applied to study the interaction between the protein of papaya eIFiso4E and PRSV-vpg.The results showed that the papaya eIFiso4E and PRSV-vpg interacted in the nucleus.The novel BiFC expression vectors are promising for a wide application for protein-protein interactions in plants.

关 键 词：分子克隆 Nimble Cloning 双分子荧光互补(BiFC) 蛋白互作 

分 类 号：S66[农业科学—果树学]