| 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. |
| 关键词: microRNA 水稻 基因克隆 Os-miR390前体基因 |
| DOI： |
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| 基金项目:扬州大学博士启动基金项目（0288900015230） |
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| Cloning and constructing of Os-miR390 precursor gene and transforming it into rice |
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| Abstract: |
| 【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 T0 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. |
| Key words: microRNA rice gene cloning Os-miR390 precursor gene |
