水稻泛素连接酶基因OsHUB2实时荧光定量PCR方法的建立

何炜; 朱永生; 连玲; 周平; 蔡秋华; 王颖姮; 谢鸿光; 张建福; 谢华安

doi:10.19303/j.issn.1008-0384.2014.07.007

水稻泛素连接酶基因OsHUB2实时荧光定量PCR方法的建立

doi: 10.19303/j.issn.1008-0384.2014.07.007

何炜^1,2,3,4,5,
朱永生^1,2,3,4,5,
连玲^1,2,3,4,5,
周平⁶,
蔡秋华^1,2,3,4,5,
王颖姮^1,2,3,4,5,
谢鸿光^1,2,3,4,5,
张建福^1,2,3,4,5,
谢华安^1,2,3,4,5

1. 福建省农业科学院水稻研究所，福建福州 350019;
2. 农业部华南杂交水稻种质创新与分子育种重点实验室/福州国家水稻改良分中心/福建省作物分子育种工程实验室/福建省水稻分子育种重点实验室，福建福州 350003;
3. 福建省作物种质创新与分子育种省部共建国家重点实验室培育基地，福建福州 350003;
4. 水稻国家工程实验室，福建福州 350003;
5. 杂交水稻国家重点实验室华南基地，福建福州 350003;
6. 福建省农业科学院果树研究所,福建福州,350013

基金项目:

国家“863”计划项目 (2014AA10A603、2014AA10A604)

福建省自然科学基金项目 (2011J01108)

详细信息

作者简介:
何炜 (1984-), 女, 硕士, 助理研究员, 主要从事水稻分子生物学研究 (E-mail:rosemary063@126.com);朱永生 (1982-), 男, 硕士, 助理研究员, 主要从事水稻遗传育种研究 (E-mail:zysfaas@qq.com);张建福 (1971-), 男, 博士, 研究员, 主要从事水稻分子生物学与分子育种研究 (E-mail:jianfzhang@163.com);谢华安 (1941-), 男, 研究员, 中国科学院院士, 主要从事水稻遗传育种研究 (E-mail:huaanxie@163.com)

中图分类号: S511
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- 被引次数: 0
出版历程
- 收稿日期: 2014-03-10
- 刊出日期: 2014-07-18

RT-PCR Analysis for Ubiquitin Ligase,OsHUB2,from Rice

HE Wei^1,2,3,4,5,
ZHU Yong-sheng^1,2,3,4,5,
LIAN Ling^1,2,3,4,5,
ZHOU Ping⁶,
CAI Qiu-hua^1,2,3,4,5,
WANG Ying-heng^1,2,3,4,5,
XIE Hong-guang^1,2,3,4,5,
ZHANG Jian-fu^1,2,3,4,5,
XIE Hua-an^1,2,3,4,5

1. Rice Research Institute,Fujian Academy of Agricultural Sciences;
2. Key Laboratory of Germplasm Innovation and Molecular Breeding of Hybrid Rice for South China,Ministry of Agriculture,P.R China/Fuzhou branch,National Rice Improvement Center of China/Fujian Engineering Laboratory of Crop Molecular Breeding/Fujian Key Laboratory of Rice Molecular Breeding;
3. Incubator of National Key Laboratory of Fujian Germplasm Innovation and Molecular Breeding between Fujian and Ministry of Sciences & Technology;
4. National Rice Engineering Laboratory of China;
5. South China Bases of National Key Laboratory of Hybrid Rice for China;
6. Fruit Research Institute,Fujian Academy of Agricultural Sciences

摘要

摘要: 在已获得水稻泛素连接酶基因OsHUB2过表达转基因植株的基础上, 根据OsHUB2基因序列设计3对引物, 分别为OsHUB2-1、OsHUB2-2、OsHUB2-3, 通过RT-PCR筛选条带特异单一的引物, real time PCR (qRT-PCR) 方法验证最适合检测OsHUB2的反应条件。结果表明OsHUB2-F3R3扩增的PCR产物特异性最强, 扩增曲线及熔解曲线等指标都在理想范围内, 同时该引物能很好地区分对照及转基因材料, 进而筛选出OsHUB2基因上调表达的转基因植株。Realtime体系的建立与优化为今后进一步研究水稻泛素连接酶基因OsHUB2的功能奠定了基础。
- 水稻 /
- 泛素连接酶基因 /
- 荧光定量PCR
Abstract: Based on the previously obtained overexpression transgenic plants with the ubiquitin ligase, OsHUB2, from rice, expression lever of the enzyme was studied.Three pairs of primers, OsHUB2-F1R1, OsHUB2-F2R2 and OsHUB2-F3R3, were designed using the Primer Premier 5.0according to the sequence of the enzyme from GeneBank.Subsequently, RT-PCR was conducted to select the best primer pairs with a specific and single amplification product.The results showed OsHUB2-F3R3 to be desirable on both and functions.The analytical procedure was thus considered of use in future studies on the rice ubiquitin ligase.
- rice /
- ubiquitin ligase OsHUB2 /
- RT-PCR (qRT-PCR)

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参考文献(13)

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