橡胶树胶乳C-乳清双向电泳样品制备技术的建立及优化

doi:10.3969/j.issn.1009-7791.2012.03.004

亚热带植物科学 ›› 2012, Vol. 41 ›› Issue (03): 16-20.DOI: 10.3969/j.issn.1009-7791.2012.03.004

橡胶树胶乳C-乳清双向电泳样品制备技术的建立及优化

戚继艳,蓝基贤,秦云霞,龙翔宇

1.中国热带农业科学院橡胶研究所，农业部橡胶树生物学重点开放实验室，海南儋州 571737；2.海南大学农学院，海南儋州 571737

收稿日期:2012-05-16 修回日期:2012-05-17 出版日期:2012-09-10 发布日期:2012-09-10
通讯作者: 龙翔宇
作者简介:戚继艳(1976-)，女，黑龙江肇东人，助理研究员，硕士，从事橡胶树产排胶机理研究。
基金资助:
农业部基本科研业务费[YWFZX2010-5(N)、RC201201]、海南省自然科学基金(310094、808187)

Optimization of Protein Preparation Suitable for 2-DE of Hevea brasiliensis C-serum

QI Ji-yan,LAN Ji-xian,QIN Yun-xia,LONG Xiang-yu

1.Key Laboratory of Rubber Biology, Ministry of Agriculture, Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou 571737, Hainan China; 2.Agricultural College of Hainan University, Danzhou, Hainan 571737, Hainan China

Received:2012-05-16 Revised:2012-05-17 Online:2012-09-10 Published:2012-09-10
Contact: Long Xiang-yu

摘要/Abstract

摘要： 在TCA/丙酮沉淀法的基础上，用乙醇和乙醚洗涤（称双乙法）沉淀获得橡胶树胶乳C-乳清蛋白样品，并在样品溶解时通过超声助溶进一步提高样品的溶解度。结果显示，利用改良后的样品制备方法（TCA/丙酮+双乙法+超声处理）所获得的C-乳清蛋白样品的溶解性大幅增加，蛋白得率高达8.47 mg/mL C-乳清，为传统TCA/丙酮法的1.6倍；同时降低了盐离子浓度，等电聚焦时升压顺利、聚焦完全，双向电泳的图谱质量明显改善，在银染2D胶上可获得1 206个蛋白点，为传统TCA/丙酮法的2.7倍。本文对胶乳C-乳清蛋白样品制备方法进行深入优化，获得了重现性较好的高质量双向电泳蛋白图谱，促进了橡胶树蛋白质组学研究。

关键词: 橡胶树C-乳清, 双向电泳, TCA/丙酮沉淀法, 双乙法, 超声助溶

Abstract: The C-serum is a major component of the latex and a key material for proteinomics of Hevea brasiliensis. Using the 2-D electrophoresis, there are some disadvantages with protein extraction from C-serum, such as unsatisfied solubility, higher ions concentrations and too many impurities. In this paper, the alcohol-ether method and sonication were used to further remove interfering substances and solubilized protein based on the traditional method of TCA/acetone. Comparing the traditional method of extraction, the results showed that the new method improved the solubility, ions concentration and purity of protein samples from C-serum. The optimized methods for extraction of protein will be extremely useful for the proteomic research of C-serum form H. brasiliensis latex.

Key words: rubber C-serum, 2-D electrophoresis, TCA precipitation, alcohol-ether, sonication solubilization

中图分类号:

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戚继艳,蓝基贤,秦云霞,龙翔宇. 橡胶树胶乳C-乳清双向电泳样品制备技术的建立及优化[J]. 亚热带植物科学, 2012, 41(03): 16-20.

QI Ji-yan,LAN Ji-xian,QIN Yun-xia,LONG Xiang-yu. Optimization of Protein Preparation Suitable for 2-DE of Hevea brasiliensis C-serum[J]. Subtropical Plant Science, 2012, 41(03): 16-20.

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橡胶树胶乳C-乳清双向电泳样品制备技术的建立及优化

Optimization of Protein Preparation Suitable for 2-DE of Hevea brasiliensis C-serum

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