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中国生物工程杂志

China Biotechnology
China Biotechnology  2011, Vol. 31 Issue (01): 51-55    DOI:
    
Comparison of the Extraction Effects of Rice Seedling Apoplast Proteins Using Different Extraction Buffers
ZHOU Lu, LIU Jin-yuan
Laboratory of Molecular Biology and MOE Laboratory of Protein Science, School of Life Science, Tsinghua University, Beijing 100084, China
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Abstract  

To date, low extraction efficiency and cytoplasmic contamination is a great problem that apoplast protein extraction faces. To effectively extract apoplast proteins, using 12 day-old 93-11 rice seedlings, the extraction efficiency and cytoplasmic contamination ratio of the apoplast protein extracts by three different extraction buffers (Buffer A, Buffer B and Buffer C) were compared. The three buffers all contain 0.1mol/L Tris-HCl pH 7.6 and 1mmol/L PMSF, however, Buffer A also contains 0.2mol/L KCl ; Buffer B also contains 0.2mol/L CaCl2; and Buffer C also contains 0.1mol/L KCl and 0.1mol/L CaCl2. The results showed that the protein yield of buffer A reached (0.49±0.07)mg/g FW(leaf)and (0.83±0.06)mg/g FW (root), respectively, 122.7% (leaf) and 102.4% (root) greater than Buffer B, and 53.1% (leaf) and 59.6% (root) greater than Buffer C. The glucose-6-phosphate dehydrogenase (G6PDH) enzyme assay showed that the cytoplasmic contamination ratios of these apoplast protein extracts were very low, below 1%, which could be considered as negligible contamination. Therefore, by optimizing the extraction buffer, an effective extraction method for apoplast proteins was established, which could be applied to the plant apoplast proteomic studies.



Key wordsRice seedling      Apoplast proteins      Extraction buffer      Condition optimization     
Received: 19 October 2010      Published: 25 January 2011
ZTFLH:  Q946.1  
Cite this article:

ZHOU Lu, LIU Jin-yuan. Comparison of the Extraction Effects of Rice Seedling Apoplast Proteins Using Different Extraction Buffers. China Biotechnology, 2011, 31(01): 51-55.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2011/V31/I01/51


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