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ptc-miR801人工microRNA表达载体构建及功能初步研究     被引量:1

Construction of Expression Vector of ptc-miR801 Gene and Function Analysis

文献类型:期刊文献

中文题名:ptc-miR801人工microRNA表达载体构建及功能初步研究

英文题名:Construction of Expression Vector of ptc-miR801 Gene and Function Analysis

作者:杨惠琴[1,2] 蒋晶[1] 刘明英[1] 乔桂荣[1] 姜彦成[2] 卓仁英[1]

第一作者:杨惠琴

机构:[1]中国林业科学研究院亚热带林业研究所;[2]新疆大学生命科学与技术学院

年份:2013

卷号:33

期号:5

起止页码:599-604

中文期刊名:植物研究

外文期刊名:Bulletin of Botanical Research

收录:CSTPCD;;北大核心:【北大核心2011】;CSCD:【CSCD2013_2014】;

基金:浙江省科技厅重大专项(2010C12010)"重要生态经济树种转基因平台建立及耐盐转基因研究"

语种:中文

中文关键词:miRNA;表达载体;拟南芥;耐盐

外文关键词:miRNA ;expression vector;Arabidop.sis thaliana ;salt stress

分类号:S287

摘要:在构建盐胁迫下青杨microRNA文库中发现了ptc-miR801,为探索植物在盐胁迫条件下ptc-miR801参与胁迫应答的机制,本实验构建了植物表达载体pCAM2300-ami801,经根癌农杆菌EHA105介导、花序侵染法获得拟南芥转基因植株。RT-PCR半定量结果显示ptc-miR801可以在转基因拟南芥中超表达且NaCl胁迫下ptc-miRNA801转基因植株种子萌发率和根长显著高于野生型,说明ptc-miR801超表达增强了转基因拟南芥耐盐性。该试验为进一步研究miR801在杨树胁迫应答机制中的作用奠定基础。
ptc-miR801 was identified from P. cathayana by microRNA library sequence, and it had been confirmed that ptc-miRS01 was induced by salt stress. The putative targets were NAC-domain protein and Scarecrow-like gene, which were shown to play important roles in response to salt stress. To investigate the function of ptc-miR801, the artificial miR801 plant expression vector was constructed and transformed into Arabidopsis thaliana. Transformed plants were confirmed by PCR and RT-PCR. The results showed that amiRNA transcript level was markedly improved in transgenic plant compared with non-transgenic plants and the overexpression of ptc-miR801 could enhance the salt tolerance of transgenic plants. Our results would help to clarify the molecular mechanism of P. cathayana under salt stress.

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