Optimization of Two Dimensional Electrophoresis System of Sugar Beet

doi:10.7668/hbnxb.2017.05.017

Abstract

Abstract: In order to obtain the best extraction method of protein in two-dimensional electrophoresis of beet leaves and establish the best two-dimensional electrophoresis system. It took beet leaves as material. To study the effects of the extraction methods of total protein,the pH value of IPG strip,and the corresponding amount of sample on the results of two-dimensional electrophoresis of sugar beet leaves through comparative analysis.The results showed that TCA/acetone extraction method was better than the phenol extraction method and soluble protein extraction method. Comparing IPG strip of pH 4-7 with pH 5-8 in two-dimensional electrophoresis:Protein spots with pH 4-7 were clear and distributed uniform in the spectra,no protein concentrated.It was more suitable for proteomics analysis of sugar beet leaves.Using the 7 cm linear phase IPG strip,protein spots with 150 μg sample volume increased obviously and was better. Using the 17 cm strip of pH 4-7 with 300 μg sample volume protein spots were more clear.The study provided the theoretical basis and reference for the study of sugar beet proteomics.

Key words: Sugar beet, Protein extraction method, Two-dimensional electrophoresis, System optimization

CLC Number:

S566.3

ZHANG Hui, BAI Chen, WANG Huazhong, ZHANG Huizhong, LI Xiaodong, FU Zengjuan, ZHAO Shangmin, E Yuanyuan, ZHANG Ziqiang, WANG Liang. Optimization of Two Dimensional Electrophoresis System of Sugar Beet[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(5): 112-116. doi: 10.7668/hbnxb.2017.05.017.

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