一个水稻小热休克蛋白基因的克隆和鉴定
收稿日期: 2016-03-30
修回日期: 2016-04-29
网络出版日期: 2016-11-10
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
Received date: 2016-03-30
Revised date: 2016-04-29
Online published: 2016-11-10
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。
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 . DOI: 10.16819/j.1001-7216.2016.6056
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
| [1] | Vierling E.The roles of heat shock proteins in plants.Annu Rev Plant Physiol Plant Mol Biol, 2003, 42(4): 579-620. |
| [2] | Wang W, Vinocur B, Shoseyov O, et al.Role of plant heat-shock proteins and molecular chaperones in the abiotic stress response.Trends Plant Sci, 2004, 9(5): 244-252. |
| [3] | Fang X, Chen J, Dai L, et al.Proteomic dissection of plant responses to various pathogens.Proteomics, 2015, 15(9): 1525-1543. |
| [4] | Li J, Xiang C Y, Yang J, et al.Interaction of HSP20 with a viral RdRp changes its sub-cellular localization and distribution pattern in plants.Sci Rep, 2015, 5: 14016. |
| [5] | Sun W, van Montagu M, Verbruggen N. Small heat shock proteins and stress tolerance in plants.Biochim Biophys Acta, 2002, 1577: 1-9. |
| [6] | Maimbo M, Ohnishi K, Hikichi Y, et al.Induction of a Small Heat Shock Protein and its functional roles in Nicotiana plants in the defense response against Ralstonia solanacearum. Plant Physiol, 2007, 145: 1588-1599. |
| [7] | Haslbeck M, Franzmann T, Weinfurtner D, et al.Some like it hot: The structure and function of small heat shock proteins.Nat Struct Mol Biol, 2005, 12: 842-846. |
| [8] | Scharf K D, Siddique M, Vierling E.The expanding family of Arabidopsis thaliana small heat stress proteins and a new family of proteins containing alpha-crystallin domains (Acd proteins). Cell Stress & Chaperon, 2001, 6(3): 225-237. |
| [9] | 羊健, 张恒木, 陈剑平, 等. 水稻黑条矮缩病毒p8蛋白的原核表达、抗血清制备及其特性. 植物保护学报, 2007, 34(3): 252-256. |
| [9] | Yang J, Zhang H M, Chen J P, et al.Prekaryotic expression antiserum preparation and some properties of p8 protein of rice black-streaked dwarf Fijivirus.Acta Phytophyl Sin, 2007, 34(3): 252-256. (in Chinese with English abstract) |
| [10] | Lewis P O, Kumar S, Tamura K, et al.MEGA6: Molecular evolutionary genetics analysis version 6.0.Mol Biol & Evol, 2013, 30(4): 2725-2729. |
| [11] | Schwede T, Kopp J, Guex N, et al.SWISS-MODEL: An automated protein homology-modeling server.Nucleic Acids Res, 2003, 31(13): 3381-3385. |
| [12] | Yang J, Zhang F, Li J, et al.Integrative analysis of the microRNAome and transcriptome illuminates the response of susceptible rice plants to rice stripe virus.PloS One, 2015, 11(1): e0146946. |
| [13] | Schmittgen T D, Livak K J.Analyzing real-time PCR data by the comparative C(T) method.Nat Prot, 2008, 3: 1101-1108. |
| [14] | 梁国栋. 最新分子生物学实验技术, 科学出版社, 北京, 2001. |
| [14] | Liang G D.The Newest Experimental Techniques of Molecular Biology, Beijing:Science Press, 2001. (in Chinese) |
| [15] | Holt S E, Aisner D L, Baur J, et al.Functional requirement of p23 and Hsp90 in telomerase complexes.Genes Dev, 1999, 13(7): 817-826. |
| [16] | Suzuki T C, Krawitz D C, Vierling E.The chloroplast small heat-shock protein oligomer is not phosphorylated and does not dissociate during heat stress in vivo.Plant Physiol, 1998, 116: 1151-1161. |
| [17] | Waters E R.The evolution, function, structure, and expression of the plant sHSPs.J Exp Bot, 2013, 64: 391-403. |
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