甜瓜N-乙酰氨基葡萄糖基转移酶基因(CmGnT)RNA干扰载体的构建  

Construction of RNA Interference Vector of CmGnT Gene in Melon

作　　者：张瑞腾 付秋实[2] 马太光 郭秀霞[1] 李灵芝[1] 郭仰东[3] 王怀松[2] 李海平[1] ZHANG Rui-teng;FU Qiu-shi;MA Tai-guang;GUO Xiu-xia;LI Ling-zhi;GUO Yang-dong;WANG Huai-song;LI Hai-ping(College of Horticulture,Shanxi Agricultural University,Taigu 030801,Shanxi,China;Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences,Beijing 100081,China;College of Horticulture,China Agricultural University,Beijing 100193,China)

机构地区：[1]山西农业大学园艺学院,山西太谷030801 [2]中国农业科学院蔬菜花卉研究所,北京100081 [3]中国农业大学园艺学院,北京100193

出　　处：《中国蔬菜》2017年第8期26-30,共5页China Vegetables

基　　金：国家自然科学基金项目(31471895;31201642);现代农业产业技术体系建设专项资金项目(CARS-26-07);中国农业科学院科技创新工程项目

摘　　要：为探讨甜瓜N-乙酰氨基葡萄糖基转移酶基因(CmGnT)的功能,选取甜瓜CmGnT基因保守序列,利用特异性引物PCR扩增得到一段238 bp的干扰片段。利用Swa I单酶切将正向片段亚克隆到质粒载体p FGC1008的35S启动子与GUS之间,利用Spe I单酶切将反向片段亚克隆到质粒载体p FGC1008的GUS与终止子之间,构建了RNAi表达载体p FGC1008-CmGnT。通过冻融法将其转入根癌农杆菌C58中,菌液PCR验证表明RNAi表达载体p FGC1008-CmGnT构建成功并成功转入农杆菌。为进一步研究甜瓜CmGnT基因功能及甜瓜分子育种奠定重要基础。In order to explore the functions of CmGnT gene,the paper selected conserved sequence ofCmGnT gene in melon,and amplified the interference fragment(238bp)by PCR with the specific primers.The forward fragment was subcloned into the plasmid vector pFGC1008between the35S promoter and GUS bySwa I single digestion,and the reverse fragment was subcloned into the plasmid vector pFGC1008between theGUS and the terminator by Spe I single digestion,and the RNAi expression vector pFGC1008-CmGnT wasconstructed.The recombinant plasmid pFGC1008-CmGnT was transformed into Agrobacterium tumefaciens(C58)by freeze-thaw method,and the results of PCR confirmed that the vector(pFGC1008-CmGnT)wassuccessfully constructed and transferred to C58.This study has laied an important basis for further exploring thefunction of CmGnT gene and molecular breeding of melon.

关 键 词：甜瓜 CmGnT基因 RNA干扰 载体构建 

分 类 号：S652[农业科学—果树学]