甜菜BvPCNA基因RNAi载体构建OA
Construction of RNAi Vector of BvPCNA Gene in Sugar Beet
[目的]增殖细胞核抗原(Proliferating cell nuclear antigen,PCNA)在植物中广泛存在,但是其在植物中的功能研究较少.本研究可以为BvPCNA-like基因的功能分析奠定基础.[方法]课题组前期在甜菜基因组中克隆获得一个基因位点,命名为BvPCNA-like基因,本研究以BvPCNA-like基因的克隆载体为初始材料,选取适宜的靶序列,利用植物表达载体pFGC5941 作为基础载体,使靶序列上游启动子为35 S,植物筛选标记为抗除草剂基因BlpR,设计靶序列特异性引物,采取PCR-酶切-连接方法,构建BvPCNA-RNAi载体.[结果]通过RCR鉴定表明已获得了预期目标载体,插入片段由正向靶序列BvPCNA-F、中间片段、反向靶序列BvPCNA-R组成,靶序列长度为320 bp.[结论]本研究所得载体适用于对双子叶植物进行遗传转化,可以作为BvPCNA-like基因理论研究或应用研究的载体材料.
[Objective]Proliferating cell nuclear antigen(PCNA)is widely existed in plants,but its function in plants has been rarely studied.This study could lay the foundation for the functional analysis of BvPCNA-like genes.[Methods]A gene site in the sugar beet genome had been isolated in the previous study,and named as BvPCNA-like gene.In this study,the cloning vector of BvPCNA-like gene was used as the initial material to select the appropriate target sequence.The plant expression vector pFGC5941 was used as the basic vector to make sure the upstream promoter of target sequence was 35 S,and the plant screening marker is the herbicide-resistant gene BlpR.Target-specific primers were designed and the BvPCNA-RNAi vector was constructed by PCR-enzyme digestion-linkage reactions.[Results]The PCR identification showed that the expected vector had been obtained,which was composed of forward target BvPCNA-F,intermediate segment,and reverse target BvPCNA-R,with a target sequence length of 320 bp.[Conclusion]The vector obtained in this study can be used for genetic transformation of dicotyledonous plants,and as a vector material for theoretical research or application research of BvPCNA-like gene.
王希;张西宁;陶艳昵;张琦;李志烨;赵春雷
陇东学院生命科学与技术学院,甘肃庆阳 745000||甘肃省陇东生物资源保护利用与生态修复重点实验室,甘肃庆阳 745000||黑龙江省普通高等学校甜菜遗传育种重点实验室/黑龙江省甜菜工程技术研究中心,哈尔滨 150080||国家糖料改良中心/中国农业科学院北方糖料作物资源与利用重点开放实验室,哈尔滨 150080黑龙江大学现代农业与生态环境学院/中国农业科学院甜菜研究所,哈尔滨 150080||黑龙江省普通高等学校甜菜遗传育种重点实验室/黑龙江省甜菜工程技术研究中心,哈尔滨 150080||国家糖料改良中心/中国农业科学院北方糖料作物资源与利用重点开放实验室,哈尔滨 150080
农业科学
甜菜BvPCNA-likeRNAi载体构建
sugar beetBvPCNA-likeRNAivector construction
《中国糖料》 2024 (002)
9-16 / 8
博士后研究人员落户黑龙江科研启动资助金"甜菜分子标记开发与重要性状相关标记的筛选"(LBH-Q18108);2017年度黑龙江省省属高等学校基本科研业务费科技创新类重点项目"甜菜分子标记位点库的深度挖掘与候选SSR标记开发"(KJCXZD201713);财政部和农业农村部国家现代农业产业技术体系(糖料)建设项目"甜菜种质资源收集与评价"(CARS-170102)资助.
评论