Two Dimensional Electrophoresis of Optimization and Extraction of Proteome Research of Maize

doi:10.7668/hbnxb.2016.04.003

Abstract

Abstract: In order to establish a set of two dimensional gel electrophoresis technique for maize,the effects of three kinds of total protein extraction methods on the results of two dimensional electrophoresis of maize were studied.Meanwhile,the protein sample volume,condition of isoelectric focusing and the density of polyacrylamide gel were explored and optimized.The results showed that TCA-acetone for total protein extraction was simple and convenient compared with the method of phenol extraction and phosphate Buffer (PB) solution method.The 2-D maps had abundant protein spots and more clear background,this study established a suitable proteomics methods for forage maize.Meantime,it screened out the first dimension electrophoresis was performed on the sample 800 μg and focus conditions 20 000 V/h;The second electrophoresis was performed on 12.5% polyacrylamide gels,it could be obtained two-dimensional electrophoresis map at 17 cm of background low and higher resolution.Optimization system would be suitable for two dimensional electrophoresis of total protein of forage maize leaves,the method can be applied to corn leaf proteome analysis of sample extraction.The useful information can be provided for the proteomics research of maize.

Key words: Maize, Leaf, Protein extraction, Two-dimensional electrophoresis (2-D)

CLC Number:

S513.03

DONG Jing, LU Xiaoping, LÜ Ersuo, XUE Chunlei, LI Junwei. Two Dimensional Electrophoresis of Optimization and Extraction of Proteome Research of Maize[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2016, 31(4): 13-18. doi: 10.7668/hbnxb.2016.04.003.

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