| 拟南芥中与植物生长和耐寒相关的miRNA表达载体的构建 |
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| 引用本文： | 蔡杰,杨成龙,段瑞军,郭建春.拟南芥中与植物生长和耐寒相关的miRNA表达载体的构建[J].广西农业生物科学,2010(4):804-808. |
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| 作者姓名： | 蔡杰 杨成龙 段瑞军 郭建春 |
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| 作者单位： | [1]海南大学农学院,海口570228 [2]中国热带农业科学院生物技术研究所,热带作物生物技术国家重点实验室,海口571101 |
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| 基金项目： | 国家自然科学基金（30860151）; 现代木薯农业产业技术体系（nycytx-17）; 中央级公益性科研院所基本科研项目（ITBBZD0721）共同资助 |
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| 摘 要： | 本研究通过PCR方法从拟南芥基因组序列中分离了3个理论推断与耐寒相关的miRNA片段（miRNA402,miRNA319和miRNA393）和1个与植物生长相关的片段miRNA172。利用重叠延伸PCR技术将miRNA172小分子,以及3个与耐寒相关的miRNA402,miRNA319和miRNA393小分子串连在一起分别导入植物表达载体pVKH-35S-GUS-pA,取代表达载体中的GUS基因,构建了pVKH-35S-mir172-pA和pVKH-35S-mir31＋402＋393-pA融合表达载体。经PCR和双酶切及测序鉴定,pVKH-35S-mir172-pA和pVKH-35S-mir319＋402＋393-pA构建成功,为后续利用基因工程手段,miRNA转化木薯,提高木薯生长和木薯耐寒相关研究奠定基础。
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| 关 键 词： | miRNA 表达载体 重叠延伸PCR技术 双酶切 |
Construction of miRNA Expression Vectors Correlative with Plant Growth and Cold-resistant in Arabidopsis |
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| Cai Jie,Yang Chenglong,Duan Ruijun,Guo Jianchun.Construction of miRNA Expression Vectors Correlative with Plant Growth and Cold-resistant in Arabidopsis[J].Journal of Guangxi Agricultural and Biological Science,2010(4):804-808. |
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| Authors: | Cai Jie Yang Chenglong Duan Ruijun Guo Jianchun |
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| Institution: | 1 College of Agriculture,Hainan University,Haikou,570228;2 Key Laboratory of Tropical Crops Biotechnology Ministry of Agriculture,Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Sciences,Haikou,571101 |
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| Abstract: | In this article,we have isolated one small molecule miRNA172 related with plant growth,3 cold-resistant related miRNA402,miRNA319 and miRNA393 from Arabidopsis genomic DNA by PCR method.Then miRNA172 and the fused miRNAs with miRNA402,miRNA319 and miRNA393 have been replaced the GUS gene in pVKH-35S-GUS-pA by splicing overlap extension PCR and constructed plant botanic expression vectors of pVKH-35S-mir172-pA and pVKH-35S-mir319＋402＋393-pA,respectively.The expressing vectors of pVKH-35S-mir172-pA,pVKH-35S-mir319＋402＋393-pA have been identified to be successfully constructed by PCR,restriction enzyme digestion and sequence analysis.The result would be the preliminary work for use of miRNAs to improve the cassava growth and cold tolerance by gene engineering method. |
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| Keywords: | miRNA Expressing vector Splicing overlap extension PCR Restriction enzyme digestion |
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