Effective protein extraction protocol for proteomics studies of Jerusalem artichoke leaves
文献类型：期刊论文
| 作者 | Zhang, Meide1; Shen, Shihua
|
| 刊名 | JOURNAL OF SEPARATION SCIENCE
 |
| 出版日期 | 2013 |
| 卷号 | 36期号:13页码:2203-2209 |
| 关键词 | 2DE Jerusalem artichoke Protein extraction Proteomics |
| ISSN号 | 1615-9306 |
| DOI | 10.1002/jssc.201300199 |
| 文献子类 | Article |
| 英文摘要 | Protein extraction is a crucial step for proteomics studies. To establish an effective protein extraction protocol suitable for two-dimensional electrophoresis (2DE) analysis in Jerusalem artichoke (Helianthus tuberosus L.), three different protein extraction methods-trichloroacetic acid/acetone, Mg/NP-40, and phenol/ammonium acetate-were evaluated using Jerusalem artichoke leaves as source materials. Of the three methods, trichloroacetic acid/acetone yielded the best protein separation pattern and highest number of protein spots in 2DE analysis. Proteins highly abundant in leaves, such as Rubisco, are typically problematic during leaf 2DE analysis, however, and this disadvantage was evident using trichloroacetic acid/acetone. To reduce the influence of abundant proteins on the detection of low-abundance proteins, we optimized the trichloroacetic acid/acetone method by incorporating a PEG fractionation approach. After optimization, 363 additional (36.2%) protein spots were detected on the 2DE gel. Our results suggest that trichloroacetic acid/acetone method is a better protein extraction technique than Mg/NP-40 and phenol/ammonium acetate in Jerusalem artichoke leaf 2DE analysis, and that trichloroacetic acid/acetone method combined with PEG fractionation procedure is the most effective approach for leaf 2DE analysis of Jerusalem artichoke. |
| 学科主题 | Chemistry, Analytical |
| 出版地 | WEINHEIM |
| 电子版国际标准刊号 | 1615-9314 |
| WOS关键词 | HELIANTHUS-TUBEROSUS ; 2-DIMENSIONAL ELECTROPHORESIS ; MASS-SPECTROMETRY ; PLANT PROTEOME ; SOLUBILIZATION ; FRACTIONATION ; EXPRESSION ; ETHANOL ; TISSUE ; INULIN |
| WOS研究方向 | Science Citation Index Expanded (SCI-EXPANDED) |
| 语种 | 英语 |
| WOS记录号 | WOS:000322567300019 |
| 出版者 | WILEY-V C H VERLAG GMBH |
| 资助机构 | Research and Development Projects in Hubei Province |
| 源URL | [http://ir.ibcas.ac.cn/handle/2S10CLM1/28095]  |
| 专题 | 中科院北方资源植物重点实验室 |
| 作者单位 | 1.Chinese Acad Sci, Inst Bot, Beijing, Peoples R China 2.Hubei Acad Agr Sci, Inst Chinese Herbal Med, Enshi 44500, Peoples R China |
| 推荐引用方式 GB/T 7714 | Zhang, Meide,Shen, Shihua. Effective protein extraction protocol for proteomics studies of Jerusalem artichoke leaves[J]. JOURNAL OF SEPARATION SCIENCE,2013,36(13):2203-2209. |
| APA | Zhang, Meide,&Shen, Shihua.(2013).Effective protein extraction protocol for proteomics studies of Jerusalem artichoke leaves.JOURNAL OF SEPARATION SCIENCE,36(13),2203-2209. |
| MLA | Zhang, Meide,et al."Effective protein extraction protocol for proteomics studies of Jerusalem artichoke leaves".JOURNAL OF SEPARATION SCIENCE 36.13(2013):2203-2209. |
入库方式： OAI收割
来源：植物研究所
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