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Expression and activity analysis of beta Gallinacin-3 in Arabidopsis

文献类型： 外文期刊

作者： Jin, Libo 1 ; Wang, Yunpeng 1 ; Xu, Nuo 1 ; Wang, Dezhong 1 ; Liu, Xiuming 2 ; Peng, Renyi 1 ; Jiang, Chao 1 ; Li, Xiaoku 1 ;

作者机构： 1.Wen Zhou Univ, Inst Life Sci, Wen Zhou 325035, Peoples R China

2.Jilin Agr Univ, Minist Educ, Engn Res Ctr Bioreactor & Pharmaceut Dev, Changchun 130118, Jilin, Peoples R China

3.Jilin Acad Agr Sci, Changchun 130118, Jilin, Peoples R China

关键词： Antimicrobial peptide;beta Gal-3;Arabidopsis;Plant reactor;Purification

期刊名称：PROTEIN EXPRESSION AND PURIFICATION （ 影响因子：1.65； 五年影响因子：1.548 ）

ISSN： 1046-5928

年卷期： 2018 年 144 卷

页码：

收录情况： SCI

摘要： beta Gallinacin-3 (beta Gal-3) is an antimicrobial peptide with strong antibacterial activity against Escherichia colt, Staphylococcus aureus and Salmonella typhimurium. In this study, the beta Gal-3 gene was transferred into a plant genome by genetic engineering techniques. These transgenic plants can be used as feed additives to prevent poultry diseases and them might replace the antibiotics used in poultry industry. To ensure the beta Gal-3 expresses effectively in Arabidopsis seeds, the expression was driven by promoter Ppha cloned from the beta-phaseolin storage protein gene. A total of 294 transgenic lines were obtained by Agrobacterium-mediated transformation into Arabidopsis, and five transgenic lines were selected in which the expression levels of beta Gal-3 were more than 0.10% of the total soluble proteins. The transgenic lines with single locus were identified by Southern blotting. The expression of beta Gal-3 and the highest protein accumulation level (about 4.76 mg/g fresh weight with a maximum of 0.27% of total soluble proteins) was measured by Western blotting and ELISA, respectively. After ultrafiltration by centrifugation, the purity of recombinant beta Gal-3 was up to 73%. Taken together, our data showed that expression of beta Gal-3 with antimicrobial activity is possible and effective in Arabidopsis seeds. (C) 2017 Published by Elsevier Inc.

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