水稻<em>OsHAK26</em>启动子的克隆及瞬时表达分析

doi:10.13523/j.cb.20170206

中国生物工程杂志

2017, Vol. 37

Issue (2): 33-39 DOI: 10.13523/j.cb.20170206

研究报告

水稻OsHAK26启动子的克隆及瞬时表达分析

罗枫雪, 李佛生, 姚民, 徐莺

四川大学生命科学学院生物资源与生态环境教育部重点实验室成都 610064

The Cloning and Transient Expression Analysis of Promoter of OsHAK26 from Oryza sativa

LUO Feng-xue, LI Fo-sheng, YAO Min, XU Ying

Key Laboratory of Bio-resources and Eco-environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610064, China

全文: PDF(716 KB) HTML

摘要：

对水稻KT/HAK/KUP钾离子转运蛋白家族OsHAK26起始密码子上游2 064bp序列进行分析，发现该序列除了具备TATA-Box、CAAT-Box等基本启动子元件外，还含有许多发育、激素、非生物胁迫等响应元件以及KT/HAK/KUP 家族启动子普遍存在的元件。用该片段及5'端缺失的－1 473bp、－963bp、－441bp、－193bp四个片段分别取代植物瞬时表达载体pBI-221的CaMV 35S启动子区域，并利用拟南芥叶肉原生质体进行瞬时表达分析。结果表明，这五种片段都具有一定的启动活性，随着长度减小，活性下降，但缺失－963bp~－441bp之间的片段却导致活性显著回升，推断该区段含有抑制元件，缺失－441bp~－193bp之间的片段导致活性大幅下降，推断－441bp~－193bp为OsHAK26基因启动子的核心启动区域。

关键词： 启动子; 瞬时表达; 水稻; OsHAK26; 缺失突变

Abstract:

A 2 064 bp upstream sequence of translation initiation site of OsHAK26, a member of KT/HAK/KUP potassium transporters gene family in rice was analyzed and that included a number of elements in response to the development, hormones, and abiotic stresses and common promoter elements of KT/HAK/KUP family, in addition to the basic promoter elements such as TATA-Box, CAAT-Box and so on. Subsequently, the fragment together with other four different 5'-end deletion fragments (－1 473bp、－963bp、－441bp、－193bp) of the OsHAK26 gene 5'-flanking sequences were inserted into the plant transient expression vector pBI221 to replace CaMV 35S promoter respectively and the transient expression analysis was conducted in Arabidopsis mesophyll protoplast system. The result showed that the five fragments had promotor activity decreasing with the length decrease. but deletion of －963bp~－441bp had led to a significant rebound in activity, so it inferred that the section contained suppression components.－441bp~+95bp region activity suddenly surged,so the －441bp~－193bp was core region of promoter of OsHAK26.

Key words: Deletion mutation Rice Transient expression OsHAK26 Promoter

收稿日期: 2016-08-19 出版日期: 2017-02-25

ZTFLH:

Q945

基金资助:

国家自然科学基金（31270360）、国家自然科学和技术重大专项（2016ZX08010001-010）资助项目

通讯作者: 徐莺 E-mail: xuying@scu.edu.cn

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引用本文:

罗枫雪, 李佛生, 姚民, 徐莺. 水稻OsHAK26启动子的克隆及瞬时表达分析[J]. 中国生物工程杂志, 2017, 37(2): 33-39.

LUO Feng-xue, LI Fo-sheng, YAO Min, XU Ying. The Cloning and Transient Expression Analysis of Promoter of OsHAK26 from Oryza sativa. China Biotechnology, 2017, 37(2): 33-39.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20170206 或 https://manu60.magtech.com.cn/biotech/CN/Y2017/V37/I2/33

[1] Clarkson D T,Hanson J B. The mineral nutrition of higher plants.Annual Review of Plant Physiology,1980,311(1):239-298.
[2] Robert H, Ralf R M. Physiological functions of mineral micronutrients (Cu, Zn, Mn, Fe, Ni, Mo, B, Cl).Current Opinion in Plant Biology,2009,12(3):259-266.
[3] Schroeder J I,Ward J M,Gassmann W. Perspectives on the physiology and structure of inward-rectifying K⁺ channels in higher plants:biophysical implications for K⁺ uptake. Annu Rev Biophys Biomol Struct,1994,23:441-471.
[4] Gupta M,Qiu X,Wang L,et al. KT/HAK/KUP potassium transporters gene family and their whole-life cycle expression profile in rice (Oryza sativa).Molecular Genetics and Genomics, 2008,280:437-452.
[5] Zhang Z,Zhang J,Chen Y,et al. Genome-wide analysis and identification of HAK potassium transporter gene family in maize (Zea mays L.).Molecular Biology Reports,2012,39:8465-8473.
[6] Rubio F,Santa-María G E,Rodríguez-navarro A. Cloning of Arabidopsis and barley cDNAs encoding HAK potassium transporters in root and shoot cells.Physiologia Plantarum,2000,109:34-43.
[7] Yang Z,Gao Q, Sun C,et al. Molecular evolution and functional divergence of HAK potassium transporter gene family in rice (Oryza sativa L.).Journal of Genetics and Genomics,2009, 36:161-172.
[8] Yang T,Zhang S,Hu Y,et al. The role of a potassium transporter OsHAK5 in potassium acquisition and transport from roots to shoots in rice at low potassium supply levels. Plant Physiology,2014,166:945-959.
[9] Bañuelos M A,Garciadeblas B,Cubero B,et al. Inventory and functional characterization of the HAK potassium transporters of rice. Plant Physiology,2002,130:784-795.
[10] Grabov A. Plant KT/KUP/HAK potassium transporters:single family-multiple functions. Annals of Botany,2007,99:1035-1041.
[11] Amrutha R N,Sekhar P N,Varshney R K,et al. Genome-wide analysis and identification of genes related to potassium transporter families in rice (Oryza sativa L.). Plant Sci,2007, 172:708-721.
[12] 聂丽娜, 夏兰琴, 徐兆师,等. 植物基因启动子的克隆及其功能研究进展. 植物遗传资源学报, 2008, 9(3):385-391. Nie L N,Xia L Q,Xu Z S,et al. Progress on cloning and functional study of plat gene pomoters.Journal of Plant Genetic Resources,2008,9(3):385-391.
[13] Yoo S,Cho Y,Sheen J. Arabidopsis mesophyll protoplasts:a versatile cell system for transient gene expression analysis. Nature Protocols,2007,2(7):1565-1572.
[14] Lescot M,Déhais P,Thijs G,et al. PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences.Nucleic Acids Res,2002,30:325-327.
[15] Higo K,Ugawa Y,Iwamoto M,et al. Plant cis-acting regulatory DNA elements (PLACE) database. Nucleic Acids Res,1999,27:297-300.
[16] Yi W, Wei H W. Potassium transport and signaling in higher plants.Annu Rev Plant Biol,2013,64:451-476.
[17] Rogers H J,Bate N,Combe J, et al. Functional analysis of cis-regulatory elements within the promoter of the tobacco late pollen gene g10.Plant Mol Biol,2001,45:577-585.
[18] Twell D,Wing R,Yamaguchi J,et al. Isolation and expression of an anther-specific gene from tomato.Mol Gen Genet,1989,217:240-245.
[19] Chen G,Hu Q D,Luo L,et al. Rice potassium transporter OsHAK1 is essential for maintaining potassium-mediated growth and functions in salt tolerance over low and high potassium concentration ranges.Plant, Cell and Environment,2015, 38:2747-2765.
[20] Abe H,Urao T,Ito T,et al. Arabidopsis AtMYC2 (bHLH) and AtMYB2 (MYB) function as transcriptional activators in abscisic acid signaling.The Plant Cell,2003,15(1):63-78.
[21] Simpson S D,Nakashima K,Narusaka Y, et al. Two divergent novel cis-acting elements of erd1, a clpA homologous Arabidopsis gene function in induction by dehydration stress and dark-induced senescence.Plant J,2003,33:259-270.
[22] Gupta M,Qiu X H,Wang L,et al. KT/HAK/KUP potassium transporters gene family and their whole-life cycle expression profile in rice (Oryza sativa).Mol Genet Genomics,2008, 280:437-452.
[23] Aida M H,Luisa L O,Gerardo A A,et al. Functional properties and regulatory complexity of a minimal RBCS light-responsive unit activated by phytochrome, cryptochrome, and plastid signals. Plant Physiol,2002,128:1223-1233.
[24] Yamamoto Y Y,Ichida H,Matsui M,et al. Identification of plant promoter constituents by analysis of local distribution of short sequences. BMC Genomics, 2007, 8:67.
[25] Oo M,Bae H K,Nguyen T,et al. Evaluation of rice promoters conferring pollen-specific expression in a heterologous system,Arabidopsis.Plant Reproduction,2014,27:47-58.

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