一个水稻小热休克蛋白基因的克隆和鉴定

doi:10.16819/j.1001-7216.2016.6056

中国水稻科学 ›› 2016, Vol. 30 ›› Issue (6): 587-592.DOI: 10.16819/j.1001-7216.2016.6056

一个水稻小热休克蛋白基因的克隆和鉴定

项聪英^1,², 蔡年俊^1,², 李静², 羊健², 陈剑平^2,^*(), 张恒木^2,^*()

¹浙江师范大学化学与生命科学学院, 浙江金华 321004
²浙江省农业科学院病毒学与生物技术研究所, 杭州310021

收稿日期:2016-03-30 修回日期:2016-04-29 出版日期:2016-11-10 发布日期:2016-11-10
通讯作者: 陈剑平,张恒木
基金资助:
浙江省自然科学基金资助项目(LQ14-C140003)

Cloning and Characterization of a Small Heat Shock Protein (SHSP) Gene in Rice Plant

Cong-ying XIANG^1,², Nian-jun CAI^1,², Jing LI², Jian YANG², Jian-ping CHEN^2,^*(), Heng-mu ZHANG^2,^*()

¹ College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004, China
² State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control, Key Laboratory of Biotechnology in Plant Protection of MOA and Zhejiang Province, Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2016-03-30 Revised:2016-04-29 Online:2016-11-10 Published:2016-11-10
Contact: Jian-ping CHEN, Heng-mu ZHANG

摘要/Abstract

摘要：

小分子热休克蛋白(SHSP)广泛存在于各类生物体中且在生物发育和逆境响应过程中发挥重要作用。我们克隆了一个水稻小热休克蛋白基因(OsSHSP17.6),序列分析表明该基因编码区为477 bp,可编码一个含158 个氨基酸的多肽,分子量约17.6 kD。序列分析显示该类SHSP在不同的植物中是保守的,在分类上隶属于CI类。实时定量RT-PCR分析显示热激处理可显著增强该基因的转录;Western blotting分析显示该蛋白大小与预期一致,且其含量在热激处理时大幅度增加。这些结果一致表明OsSHSP17.6可能参与了水稻热应激反应,这为进一步鉴定OsSHSP17.6的功能提供了基础。

关键词: 小热休克蛋白, 热激处理, 表达模式

Abstract:

Small heat shock proteins (SHSPs) exist widely in all kinds of creatures and play an important role in both development and stress responses. A gene encoding a SHSP (OsSHSP17.6) was cloned from Oryza sativa and the sequence assembly showed the encoding region of OsSHSP17.6 was 477 bp in length, encoding a polypeptide of 158 aa with a molecular weight of 17.6 kD. Sequence analysis showed that such kind of SHSPs was conserved in different plants and belonged to the cytosolic class I (CI). The transcriptional level of OsSHSP17.6 was significantly up-regulated by heat shock with qRT-PCR. The protein appeared to be accumulated up to higher level under the condition of heat shock in western blotting assays. Taken together, these findings consistently indicated that OsSHPS17.6 could be involved into the response to heat shock, which could contribute to understanding the function of OsSHSP17.6.

Key words: small heat shock protein (SHSP), heat shock, expression pattern

中图分类号:

Q785
S511.01

项聪英, 蔡年俊, 李静, 羊健, 陈剑平, 张恒木. 一个水稻小热休克蛋白基因的克隆和鉴定[J]. 中国水稻科学, 2016, 30(6): 587-592.

Cong-ying XIANG, Nian-jun CAI, Jing LI, Jian YANG, Jian-ping CHEN, Heng-mu ZHANG. Cloning and Characterization of a Small Heat Shock Protein (SHSP) Gene in Rice Plant[J]. Chinese Journal OF Rice Science, 2016, 30(6): 587-592.

图/表 4

图1 OsSHSP17.6序列同源性(A)及保守结构域(B)分析登录号分别为:SiSHSP(XP004984667,谷子)、AtSHSP(EMT09347,粗山羊草)、BdSHSP(XP010228221,二穗短柄草)、ZmSHSP(ACG35098,玉米)、SbSHSP(KXG39718,高粱)。

Fig. 1. Sequence homology (A) and conserved domain (B) analyses of OsSHSP17.6. Accession numbers: SiSHSP (XP004984667, Setaria italica), AtSHSP (EMT09347, Aegilops tauschii), BdSHSP (XP010228221, Brachypodium distachyon ), ZmSHSP (ACG35098, Zea mays), SbSHSP (KXG39718,Sorghum bicolor).

图2 OsSHSP17.6与拟南芥中的小热休克蛋白进化关系

Fig. 2. Phylogenetic relationship of OsSHSP17.6 with SHSPs from Arabidopsis thaliana.

图3 实时定量PCR分析OsSHSP17.6基因转录水平

Fig. 3. Transcription level of OsSHSP17.6 by real-time quantitative PCR.

图4 Western blot分析OsSHSP17.6蛋白的表达水平 M-蛋白标记; 第1泳道为对照组样品; 第2泳道为热激处理样品。

Fig. 4. Translational level of OsSHSP17.6 by Western blotting. M, Protein marker; Lane 1, Control sample; Lane 2, Sample treated with heat shock.

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一个水稻小热休克蛋白基因的克隆和鉴定

Cloning and Characterization of a Small Heat Shock Protein (SHSP) Gene in Rice Plant

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