The Modified Method for Cotton Root Total Proteins Extraction and Two-dimensional Electrophoreses

doi:10.7668/hbnxb.2015.06.019

Abstract

Abstract: To explore an efficient and high resolution 2-DE procedure for cotton root proteomic analysis.Compared with three different protein extraction methods of cotton ( Gossypium hirsutum L.) root and analysis protein content,protein purity and electrophoresis.The results showed that the protein content of cotton root extracted by saturated powder was higher than that extracted by other methods and the electrophoresis strip of SDS-PAGE were clear;there were 601 clear cotton protein pots in a 2D-PAGE map.The TCA/acetone method got the highest protein quality and the lowest purity.The 2D-PAGE map was fuzzy and the number of protein pots was only 417.The loading quantity of 1 500 μg appeared to be better for 2-DE images with high definition when we used coomassie brilliant stain.In short,cotton root protein extraction method by saturated powder could get good separation effect.

Key words: Cotton root, Protein extraction, Two-dimensional electrophoresis, Proteomics

CLC Number:

S562.03

WANG Man, LIU Lian-tao, SUN Hong-chun, ZHANG Yong-jiang, SONG Shi-jia, CHEN Jing, CHENG Guo-peng, LI Cun-dong. The Modified Method for Cotton Root Total Proteins Extraction and Two-dimensional Electrophoreses[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(6): 128-133. doi: 10.7668/hbnxb.2015.06.019.

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References

[1] Rossignol M,Peltier J B,Mock H P,et al.Plant proteome analysis[J].Proteomics,2006,6(2):285-298.
[2] Sheoran I S,Ross A R,Olson D J,et al.Compatibility of plant protein extraction methods with mass spectrometry for proteome analysis[J].Plant Science,2009,176(1):99-104.
[3] Chen Q,Zhang M D,Shen S H.Comparison of protein extraction methods suitable for proteomics analysis in seedling roots of Jerusalem artichoke under salt (NaCl) stress[J].African Journal of Biotechnology,2011,10(39):7650-7657.
[4] 王雪,马骏,张桂寅,等.黄萎病菌胁迫条件下棉花叶片的蛋白质组分析[J].棉花学报,2007(4):273-278.
[5] 徐子剑,舒骁,杨亦玮,等.棉花纤维蛋白质3种提取及二维电泳方法的比较[J].中国生物化学与分子生物学报,2006,22(1):77-80.
[6] 林必博.棉花蛋白提取方法及其蛋白质组学研究进展[J].山西农业科学,2011,39(7):740-743.
[7] Mireille F,Esther P,Jamila C.Phenol extraction of proteins for portcomic studies of recalcitrant plant tissues [J].Methods in Molecular Biology,2007,355:9-14.
[8] 秦利征,赵全志,李俊周.水稻根系蛋白质提取方法的改良[J].分子植物育种,2013(4):588-594.
[9] 丁承强,马丹,王绍华,等.水稻蛋白质组双向电泳优化流程及方法[J].植物学报,2011,46(1):67-73.
[10] 郭忠军,陈全家,许春华,等.棉花叶片蛋白质组双向电泳技术的优化[J].棉花学报,2012(5):468-472.
[11] 刘丽娟,舒烈波,陈海荣,等.适用于生菜叶片蛋白质双向电泳方法的建立及初步应用[J].植物遗传资源学报,2009,10(1):106-110.
[12] 王珊珊,郝宇涵,任婧祺,等.黄瓜根系蛋白质组分析的双向电泳技术条件优化[J].沈阳农业大学学报,2012,43(2):233-236.
[13] Yao Y,Yang Y W,Liu J Y.An efficient protein preparation for proteomic analysis of developing cotton fibers by 2-DE[J].Electrophoresis,2006,27(22):4559-4569.
[14] Wang W,Scali M,Vignani R,et al.Protein extraction for two-dimensional electrophoresis from olive leaf,a plant tissue containing high levels of interfering compounds[J].Electrophoresis,2003,24(14):2369-2375.
[15] Wang S N,Lee K T,Ker C G.Leptin in hepatocellular carcinoma[J].World Journal of Gastroenterology,2010,16(46):5801-5809.
[16] Görg A,Obermaier C,Boguth G,et al.The current state of two-dimension electrophoresis with immobilized pH gradients[J].Electrophoresis,2000,21(6):1037-1053.
[17] 张根连.陆地棉叶片早衰的比较蛋白质组学研究[D].北京:中国农业科学院,2011.
[18] 孙立荣,胡盼盼,李亚平,等.棉花根总蛋白双向电泳方法的优化[J].河南大学学报:自然科学版,2014,44(3):329-333.
[19] 孙锋,王云生.盐胁迫下AM菌侵染的棉花幼苗根系蛋白质组分析[J].核农学报,2012(1):170-175.
[20] 杨玉霞.双向电泳技术在蛋白质组学研究中的应用进展[J].浙江农业科学,2011(6):1415-1416.
[21] 郭广芳,董坤,高利艳,等.小麦根蛋白提取与双向电泳方法的优化与应用[J].植物遗传资源学报,2010(2):244-248.

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