Leaf Protein Extraction Methods Suitable for SDS-PAGE Analysis in Apple

doi:10.7668/hbnxb.2009.02.016

Abstract

Abstract: In order to develop an efficient protein extraction method suitable for SDS - PAGE analysis,five protocols for protein extraction in apple leaf,trichloroacetic acid/acetone precipitation (TCA),modified Tris - HCl extraction,phenol extraction methanol/ ammonium acetate precipitation,DTT/ acetone and Tris - HCl extraction were compared.The extracts were separated by SDS - PAGE method followed by silver staining. The results showed that the modified Tris-HCl extraction presented the highest resolution of 3. 243μg/μL and the maximum bands of 41 strips with the 5μL sample volumes basing on the increased amount of PVPP and the extracting solution as well as prolonged 30 min of protein precipitation with cold acetone.Dynamic changes of proteins in leaves from different nodes in apple seedlings (Jonathan ×Golden Delicious)were analyzed with the improved Tris - HCl extraction method,six protein bands were detected with the molecular weight 71.9,60.5,52.6,41.1,35.3,18.5 kDa respectively and the bands were clear with a light background.So the improved Tris - HCl extraction was the most appropriate method for apple leaf SDS - PAGE analysis。

Key words: Apple, SDS - PAGE, Extraction

CLC Number:

S661.1

ZENG Guang-juan, LI Chun-min, ZHANG Xin-zhong, CHEN Dong-mei, ZHAO Yong-bo, DONGWen-xuan. Leaf Protein Extraction Methods Suitable for SDS-PAGE Analysis in Apple[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2009, 24(2): 75-78. doi: 10.7668/hbnxb.2009.02.016.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.7668/hbnxb.2009.02.016

http://www.hbnxb.net/EN/Y2009/V24/I2/75

[1]	JIA Chaoshuang, WANG Zhihua, WANG Wenhui. Effects of Different Shelf Temperature Combined with 1-MCP Treatment on Texture Traits of Huahong and Huayue Apples [J]. Acta Agriculturae Boreali-Sinica, 2022, 37(4): 128-140.
[2]	LU Xiaoyue, WANG Ziye, HAN Jianwei, XU Lu, ZHANG Xiaofei, HAN Jie, WANG Zhigang, LIU Yue, SUO Xiangmin, YAN Aihua. Effects of Pyroligneous Acids on the Growth and Main Characteristics of Replanted Soil of Malus angustifolia [J]. Acta Agriculturae Boreali-Sinica, 2022, 37(3): 175-185.
[3]	ZHAO Huiying, YANG Guangkai, GAO Yan, ZHANG Xiaojun, HAO Yanyan. *Cloning of Gene MdSGR2* from Apple and Construction of Plant Overexpression Vector** [J]. Acta Agriculturae Boreali-Sinica, 2022, 37(2): 42-48.
[4]	WANG Qian, CAI Huacheng, WANG Shuting, ZHANG Xiaojun, GAO Jingdong, DU Xuemei, LI Chunyan, HAO Yanyan, YANG Tingzhen. Comparision on Growth and Photosynthetic Characteristics of Fuji Apple with Two Intermediate Rootstocks under High Spindle [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(1): 108-115.
[5]	LIU Ziji, LIU Weixia, NIU Yu, YANG Yan. Effects of Grafting on Volatile Flavor Compounds in Yellow Cherry Tomato Fruit [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(S1): 176-182.
[6]	FU Jing, SUN Meng, MA Chunhui, HUANG Yonghong. Metabolomic Analysis of Four Allium Species and Their Inhibitory Effects on Botryosphaeria berengriana [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(S1): 363-370.
[7]	LI Shijun, YIN Baoying, LI Jia, LI Zhongyong, ZHANG Xueying, XU Jizhong. Changes of Ethylene, Related Enzymes and Their Genes Expression During Softening of Premature Apple Fruits [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(6): 104-109.
[8]	WANG Peng, WANG Junjie, LI Zhenbiao. *Extraction Method of Total Protein from Trichothecium roseum* Suitable for Proteomic Analysis** [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(3): 209-216.
[9]	YANG Rui, HAN Xuechun, LI Qing, CHEN Zhaorong. Screening of Pear Viral Disease Detection Methods and Detection of Pear Virus Disease in Tianjin Region [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2018, 33(5): 125-129.
[10]	ZHANG Haiyue, XIAO Yuxiong, TIAN Yike, WANG Caihong, YANG Shaolan, LI Yu, LI Yuqi. Sequence Characters Analysis of a β-Tubulin Gene and Its Promoter in Apple [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2018, 33(3): 78-85.
[11]	JIN Qin, DAI Peihong, LIU Chao, HU Qinhua, LIU Xiaodong, LI Yue. *Cloning and Expression Analysis of Four Apple MdCBF* Genes** [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(S1): 17-22.
[12]	WANG Tao, WANG Chaonan, ZHANG Hong, WEN Juanjuan, ZHANG Bin. Study on Rapid Extraction of Genomic DNA from Chinese Cabbage [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(6): 67-72.
[13]	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.
[14]	HU Jiayi, LUO Zhiwen, HE Fan, LI Xianghong, HU Fuchu, FAN Hongyan, LIU Zhixin, ZHANG Zhili. Establishment of a Triple RT-qPCR Method for the Simultaneous Detecting of PMWaV-1, PMWaV-2 and PMWaV-3 [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(2): 38-44.
[15]	ZHANG Guojun, WANG Tingting, HU Lizong, LI Shufen, GAO Wujun. Sequence Characterization, Expression, and Evolutionary Analysis of Heat Shock Transcription Factors in Apple [J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(2): 71-80.

Please choose a citation manager

Content to export