甘蔗SoInvInh2基因克隆及其在转基因拟南芥中的表达分析  

Cloning of Invertase Inhibitor Gene(SoInvInh2)in Sugarcane and Its Expression in Transgenic Arabidopsis

作　　者：牛俊奇 苗小荣[1] 王道波 黄维 杨丽涛 Niu Junqi;Miao Xiaorong;Wang Daobo;Huang Wei;Yang Litao(College of Biology and Pharmacy,Yulin Normal University,Yulin,537000;Guangxi Key Laboratory of Agricultural Resources Chemistry and Biotechnology,Yulin Normal University,Yulin,537000;College of Agriculture,Guangxi Universily,Nanning,530005)

机构地区：[1]玉林师范学院生物与制药学院,玉林537000 [2]玉林师范学院,广西农产资源化学与生物技术重点实验室,玉林537000 [3]广西大学农学院,南宁530005

出　　处：《分子植物育种》2020年第11期3587-3592,共6页Molecular Plant Breeding

基　　金：国家自然科学基金(31860403);广西自然科学基金项目(2017GXNSFBAl98023);广西高校中青年教师科研基础能力提升项目(2019KY0597);玉林师范学院校级科研项目(2018YJKY27)共同资助

摘　　要：转化酶抑制子(InvInh)是转化酶翻译后水平的一个重要调控因子,通过与转化酶结合成复合物来抑制转化酶酶活性。为了探究甘蔗SoInvInh2基因的功能特性,本研究以甘蔗品种GT28叶片总RNA为模板,采用RT-PCR技术扩增甘蔗SoInvInh2基因的编码区序列,以中间载体pMD18-T和表达载体pRI101-AN为基础,采用酶切和连接的方法,构建植物正义表达载体。通过花序轴浸染法转化拟南芥,获得转SoInvInh2基因拟南芥株系。经kan抗性筛选和PCR扩增检测,共获得12株阳性植株,转化效率为9.7%。半定量RT-PCR分析结果表明SoInvInh2基因在转基因拟南芥叶中相对表达量最高,根中次之,叶柄中最低。这将为后续解析甘蔗SoInvInh2基因的功能提供理论参考。Invertase inhibitor(InvInh)is an important regulatory factor after the post-translational of invertase,which inhibits the activity of invertase(INV)by combining with INV to form a complex.In order to explore the functional characteristics of the SoInvInh2 gene of sugarcane,the coding region of the SoInvInh2 gene was amplified by RT-PCR using the total RNA of the sugarcane GT28 leaf as a template,based on the intermediate vector p MD18-T and the plant expression vector pRI101-AN.A plant sense expression vector was constructed by restriction enzyme digestion and ligation methods.Arabidopsis thaliana was transformed by inflorescence axis dip method,and the transgenic SoInvInh2 gene Arabidopsis thaliana strain was obtained.A total of 12 positive plants were obtained by kan resistance screening and PCR amplification,and the transformation efficiency was 9.7%.The results obtained by semi-quantitative RT-PCR method further suggested that the SoInvInh2 gene had the highest relative expression in leaves,followed by roots and lowest in petiole in transformed plants.This work could be helpful for the functional analysis of the SoInvInh2 gene.

关 键 词：甘蔗(Saccharum officinarum) 转化酶抑制子 遗传转化 过表达 

分 类 号：S566.1[农业科学—作物学] Q943.2[生物学—植物学]