利用抑制差减杂交技术分离三角褐指藻在缺氮条件下上调表达的基因

唐建新，陈卓，胡晗华

doi:10.3724/SP.J.1005.2009.00865

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2009 , Vol. 31 >Issue 8: 865 - 870

DOI: https://doi.org/10.3724/SP.J.1005.2009.00865

研究报告

利用抑制差减杂交技术分离三角褐指藻在缺氮条件下上调表达的基因

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1. 中国科学院水生生物研究所淡水生态与生物技术国家重点实验室, 武汉 430072;
2. 中国科学院研究生院, 北京 100049

收稿日期: 2009-02-13

修回日期: 2009-05-25

网络出版日期: 2009-08-10

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Separation of the up-regulated genes under nitrogen starvation from Phaeodactylum tricornutum by suppression subtractive hybridization technology

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1. State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China;
2. Graduate School of the Chinese Academy of Sciences, Beijing 100049, China

Received date: 2009-02-13

Revised date: 2009-05-25

Online published: 2009-08-10

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摘要

为了阐明硅藻利用氮源的分子机制, 以三角褐指藻为材料, 利用抑制差减杂交技术, 分离鉴定了16个在缺氮诱导条件下上调表达的基因片段。其中, 与已知功能基因具有较高相似性的有7种, 都是跟氮源的吸收利用相关的。Northern blotting验证其中5个基因, 包括硝酸盐转运蛋白基因、铁氧化还原蛋白亚硝酸还原酶基因、铵盐转运蛋白基因、结合ATP盒的转运蛋白基因和嘌呤透过酶基因, 在缺氮诱导条件下转录水平有明显上调。

关键词： 抑制性差减杂交; 三角褐指藻; 缺氮

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唐建新，陈卓，胡晗华 . 利用抑制差减杂交技术分离三角褐指藻在缺氮条件下上调表达的基因[J]. 遗传, 2009 , 31(8) : 865 -870 . DOI: 10.3724/SP.J.1005.2009.00865

Abstract

Sixteen cDNA fragments of Phaeodactylum tricornutum were isolated and identified to be up-regulated under nitrogen starvation by suppression subtractive hybridization (SSH) technology in order to elucidate the molecular mecha-nism of the diatom nitrogen utilization. Seven of them have high similarity with the functional genes related to the utiliza-tion of nitrogen. Northern blotting analysis indicated that 5 genes, nitrate transporter gene (nrt), nitrite reductase gene (nir), ammonium transporter gene (amt), ATP-binding cassette transporter gene (abc), and purine permease gene (pup), were significantly up-regulated under nitrogen starvation.

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