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Analysis of Proteome Changes in Cold-acclimated Saussurea involucrata Callus by an Improved Efficient Protein Extraction Method
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- Author:
- No author available
- Journal Title:
- Acta Botanica Boreali-Occidentalia Sinica
- Issue:
- 9
- DOI:
- 10.3969/j.issn.1000-4025.2012.09.027
- Key Word:
- 雪莲;双向电泳;样品制备;蛋白质组
Abstract： 以产自西藏绵头雪莲为材料,分别采用TCA/丙酮法、尿素法和酚法,提取雪莲愈伤组织蛋白质并进行双向电泳,对蛋白产量和纯度以及电泳图谱进行比较.结果表明:(1)酚法较TCA/丙酮法和尿素法获得的蛋白质更纯,杂质少,蛋白点多且分辨率较高,在二维电泳图谱中背景清晰,横纹和纵纹较少.(2)对酚法提取的蛋白质样品进行上样量的比较结果显示,17 cm胶条500 μg上样量二维图谱的背景和蛋白点分布较好.(3)用0℃处理雪莲愈伤组织12h,利用建立的双向电泳体系分析结果发现,有33个蛋白点比常温(23℃)上调1.5倍表达,有5个蛋白点的表达下调2倍.(4)质谱鉴定结果表明,低温诱导的蛋白包括NADP-依赖型异柠檬酸脱氢酶、腺苷高半胱氨酸酶、抗性RPP8类蛋白、蛋白点gi│13129470、α-微管蛋白,分别与新陈代谢、植物防御、能量代谢、细胞的结构蛋白相关.
- [1]陈玉珍,李凤兰.低温锻炼对绵头雪莲花组织培养苗抗寒性及抗氧化酶活性的影响[J].2005,(4).
- [2]陈玉珍,卢存福.水母雪莲愈伤组织和悬浮培养细胞的抗冻性研究[J].2002,(2).
- [3]邓贵明,曾继吾,姜波,等.柑橘叶片和果皮总蛋白质提取及双向电泳体系的建立[J].2012,(1).
- [4]任丽萍,范海延,王珊珊,等.黄瓜叶片蛋白质双向电泳样品分级优化[J].2011,(8).
- [5]李红兵,康振生.适于小麦叶片蛋白质组分析的样品提取方法研究[J].2011,(3).
- [6]CHEN Yu-zhen,LI Feng-lan.Micropropagation and Callus Culture of Saussurea laniceps, an Alpine Medicinal Plant[J].2005,(1).
- [7]Prof. Dr. Volker Neuhoff,Reinhard Stamm,Hansjörg Eibl.Clear background and highly sensitive protein staining with Coomassie Blue dyes in polyacrylamide gels: A systematic analysis[J].1985,6(9).
- [8]Riederer BM,Shaw MM.Sample preparation for two-dimensional gel electrophoresis.[J].2003,3(8).
- [9]Kinney AJ,Herman EM,Helm RM,etc.Genetic modification removes an immunodominant allergen from soybean[J].2003,132(1).
- [10]A, G?rg,C, Obermaier,G, Boguth,etc.The current state of two-dimensional electrophoresis with immobilized pH gradients.[J].2000,21(6).
- [11]Nordhoff E.,Gobom J.,Giavalisco P.,etc.Extraction of proteins from plant tissues for two-dimensional electrophoresis analysis[J].2003,24(1).
- [12]Babu Raman,Jie Shen,Mark R. Marten,etc.Solubilization of Trichloroacetic Acid (TCA) Precipitated Microbial Proteins via NaOH for Two-Dimensional Electrophoresis[J].2003,2(1).
- [13]F, Cremer,C, Van de Walle.Method for extraction of proteins from green plant tissues for two-dimensional polyacrylamide gel electrophoresis.[J].1985,147(1).
- [14]M M, Bradford.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.[J].1976,72.
- [15]Zukas AA,Breksa AP.Extraction methods for analysis of Citrus leaf proteins by two-dimensional gel electrophoresis[J].2005,1078(1).
- [16]Saravanan RS,Rose JKC.A critical evaluation of sample extraction techniques for enhanced proteomic analysis of recalcitrant plant tissues[J].2004,4(9).
- [17]谭敦炎,朱建雯.雪莲的生殖生态学研究[J].1998,021 (1).