Procedure to Prepare Samples for Two-dimensional Electrophoresis of Secreted Proteins from Saccharomyces cerevisiae

doi:10.13560/j.cnki.biotech.bull.1985.2015.05.012

Abstract

Abstract: Saccraomyces cerevsiae FFC2144 was cultured in nitrogen base medium without protein. The secretory proteins of yeasts were extracted by ammonium sulfate precipitation, ultrafiltration and lyophilization-phenol extraction respectively. Extraction rates by 3 methods were calculated and the proteins were separated by two-dimensional electrophoresis. The proteins isolated were confirmed by MALDI-TOF-MS. The extraction rate by lyophilization-phenol method was 73.67% and 114 protein spots were obtained with the most protein spots and clearest electrophoretogram. Lyophilization-phenol method could be an ideal separation method for studying secretory proteomics.

Key words: Saccharomyces cerevisiae, secretory protein, two-dimensional electrophoresis, extraction method

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Du Wei, Piao Yongzhe, Huang Wei, Gu Yue, Zhao Changxin. Procedure to Prepare Samples for Two-dimensional Electrophoresis of Secreted Proteins from Saccharomyces cerevisiae[J]. Biotechnology Bulletin, 2015, 31(5): 73-77.

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Procedure to Prepare Samples for Two-dimensional Electrophoresis of Secreted Proteins from Saccharomyces cerevisiae

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[1]	WANG Zhi-wen, CHEN Hai-bo, SONG Fu-ping, GUO Shu-yuan. Identification and Analysis of Secretory Proteins in Bacillus thuringiensis [J]. Biotechnology Bulletin, 2017, 33(4): 169-176.
[2]	CHEN Fei, ZHOU Tong, WEI Yu-jia, YANG Jing, DAI Chuan-chao. Establishment of Membrane Proteomics Platform with Two-dimensional Electrophoresis for Preparing Identifying Plasma Membrane Proteins from Atractylodes lancea [J]. Biotechnology Bulletin, 2016, 32(9): 72-82.
[3]	Xia Rizhao, Liao Xiaolan. Research Progress of Agricultural Crop Active Component Extraction [J]. Biotechnology Bulletin, 2015, 31(3): 49-56.
[4]	Gu Yue, Piao Yongzhe, Du Wei, Wang Xiaoyu, Wang Chunyan. A Protein Differential Analysis of Cell Wall in Saccharomyces cerevisiae Under Different Temperatures [J]. Biotechnology Bulletin, 2015, 31(12): 200-206.
[5]	Wan Yue, Qi Jiying, Zeng Hong, Han Yang, Han Jing. Optimization of Ultrasonic Assisted Extraction of Lentinan by Response Surface Methodology [J]. Biotechnology Bulletin, 2015, 31(1): 79-85.
[6]	Liu Lin, Jia Peipei, Lu Weidong, Guo Qilin, Guo Lizhong. Differential Protein Expression Analysis of Hypsizygus marmoreus Under High Temperature Stress [J]. Biotechnology Bulletin, 2014, 0(5): 142-147.
[7]	Liu Xiangbiao, Niu Wenting, Duan Jiangyan. Influence on Wheat Seed Embryo Proteins During the Germination Period by Changed NaCl Stress [J]. Biotechnology Bulletin, 2013, 0(5): 63-68.
[8]	Wang Feifei, Wu Kunyi, Guo Ling, Cui Langjun, Ren Jing . Optimization and Construction of the Intracellular and Extracellular Proteomic Map of Lysobacter yanansis sp. nov. [J]. Biotechnology Bulletin, 2013, 0(4): 140-146.
[9]	Qi Tingting, Duan Jiangyan. UV-B Radiation Affects Protein Expression of the Wheat’s Leaf [J]. Biotechnology Bulletin, 2013, 0(3): 53-59.
[10]	Liu Xiangbiao, Duan Jiangyan. Analysis of Differentially Expressed Proteins of Embryo During Seed Germination in Wheat [J]. Biotechnology Bulletin, 2013, 0(3): 60-64.
[11]	Zhang Sheng, Zhao Zhong, Liu Zhaojun, Zhang BoyongZong Jianwei, Zhang Lingling, . Establishment and Application of Two-dimensional Gel Electrophoresis for Proteome from the Phloem of Tetraploid Robinia pseudoacacia [J]. Biotechnology Bulletin, 2013, 0(1): 116-122.