Protein extraction from rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) root for two-dimensional electrophresis |
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| Authors: | Xiaoliang Xiang Shuju Ning Xia Jiang Xiaogui Gong Renlei Zhu Lanfang Zhu Daozhi Wei |
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| Institution: | (1) Beijing Key Laboratory of Plant Resources Research and Development, Beijing Technology and Business University, Beijing, 100037, China |
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| Abstract: | An efficient protein extraction method for two-dimensional gel electrophoresis (2-DE) from plant samples is usually challenging
due to the low protein content and high level of interfering compounds. Proteomic analyses of rice (Oryza sativa L.) roots are limited by the lack of an efficient protein extraction method. To establish an effective protocol of protein
extraction suitable for 2-DE analysis in rice roots, we evaluated three protein extraction methods (trichloroacetic acid TCA]/acetone,
Mg/NP-40/TCA, and tris-base/acetone). Our results showed that the Mg/NP-40/TCA extraction method had the highest protein yield
and is the best resolution of protein separation among the three methods. The TCA/acetone method exhibited clear protein profiles
and detected more protein spots with the highest intensity in the region of high Mr (above 45 kDa) than the other methods.
However, this method was unable to detect proteins with low-Mr (less than 24.0 kDa). The Tris-base/acetone method showed the
poorest resolution of protein separation. Our results suggest that the Mg/NP-40/TCA method was the most effective among the
three methods and may provide enhanced proteomic information for rice and other crop roots. |
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