水稻Os-miR390前体基因的克隆、载体构建及转化  

Cloning and constructing of Os-miR390 precursor gene and transforming it into rice

作　　者：鲁玉柱[1] 封振[1] 边黎颖[1] 梁建生[1] 

机构地区：[1]扬州大学生物科学与技术学院,江苏扬州225009

出　　处：《西北农林科技大学学报（自然科学版）》2009年第8期87-91,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：扬州大学博士启动基金项目(0288900015230)

摘　　要：【目的】microRNA是一类长度为21~23 nt的非编码小分子RNA,对动植物的正常发育具有非常重要的调节作用。水稻Os-miR390的功能可能与生长素应答相关,拟采用转基因方法研究其生物学功能。【方法】根据水稻Os-miR390的前体RNA结构特点合理设计引物,从水稻基因组中克隆出其前体DNA片段,将其构建在玉米UbiI启动子驱动的表达载体中,并转入水稻。【结果】利用PCR成功地从水稻基因组中扩增出了Os-miR390前体基因,并将其构建在UbiI水稻组成型启动子下。经农杆菌介导,将其成功转入水稻愈伤,获得抗性幼苗,经过PCR检测,得到阳性转基因植株。【结论】T0代转基因植株表现叶片黄化的特征,待T1至T2代性状稳定后结合研究靶基因可深入研究其功能。【Objective】 MicroRNAs are a class of about 21-23 nt,non-codding,and small RNA that play important roles in the development of plants and animals.The roles of Os-miR390 may be related with Auxin response.Here transgenic methods were used to explore the function of Os-miR390 in rice.【Method】 Using the molecular cloning methods to clone,construct the precursor of Os-miR390 and transform it into rice.【Result】 The precursor DNA of Os-miR390 from rice genomic DNA was successfully amplified and constructed under the Ubiquitin Ⅰ constitutive promoter. Thereafter it was introduced into rice mediated by Agrobacterium and the positive transgenic plants were gained by PCR. [Conclusion] Although the To transgenic plants show a phenotype of yellow leaf,the roles of miR390 can be explored in-depth through combining the research of the target genes after the traits of T1 to T2 have stablized.

关 键 词：MICRORNA 水稻 基因克隆 Os-miR390前体基因 

分 类 号：Q785[生物学—分子生物学]