| 香蕉多酚氧化酶成熟蛋白的原核表达 |
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| 引用本文： | 陈 娇,孙长君,杨 昭,王朝政,李奕星,李芬芳,袁德保,谭 琳,仇厚援,陈文学,金志强.香蕉多酚氧化酶成熟蛋白的原核表达[J].热带作物学报,2015,36(12):2198-2203. |
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| 作者姓名： | 陈 娇 孙长君 杨 昭 王朝政 李奕星 李芬芳 袁德保 谭 琳 仇厚援 陈文学 金志强 |
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| 作者单位： | 中国热带农业科学院海口实验站/海南省香蕉遗传改良重点实验室;中国热带农业科学院海口实验站/海南省香蕉遗传改良重点实验室;海南大学食品学院;中国热带农业科学院海口实验站/海南省香蕉遗传改良重点实验室;中国热带农业科学院海口实验站/海南省香蕉遗传改良重点实验室;中国热带农业科学院海口实验站/海南省香蕉遗传改良重点实验室;中国热带农业科学院海口实验站/海南省香蕉遗传改良重点实验室;中国热带农业科学院海口实验站/海南省香蕉遗传改良重点实验室;海南大学食品学院;海南大学食品学院;中国热带农业科学院海口实验站/海南省香蕉遗传改良重点实验室 |
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| 基金项目： | 国家自然科学基金青年科学基金（No. 31101328）；中国热带农业科学院基本科研业务费青年拔尖人才项目（No. 1630052014005)。 |
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| 摘 要： | 笔者所在课题组从巴西香蕉中分离到1个PPO基因全长，并进行了原核表达，但结果未表达出目的蛋白。鉴于上述情况，笔者对香蕉PPO全蛋白进行转移肽分析，结果发现其N端含有转移肽，长度为47个氨基酸。切除转移肽并进行香蕉PPO成熟蛋白原核表达，SDS-PAGE电泳检测结果表明，在62 ku处有一条特异的蛋白条带，与预测大小相符；并且进行质谱分析，结果确定重组表达蛋白为香蕉PPO，初步说明转移肽对香蕉PPO体外表达的影响，为进一步研究香蕉PPO功能及在香蕉褐变生理和调控机制中的作用提供理论基础。
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| 关 键 词： | 香蕉 多酚氧化酶 转移肽 成熟蛋白 原核表达 |
Prokaryotic Expression of Polyphenol Oxidase Mature Protein from Banana (Musa acuminate) |
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| CHEN Jiao,SUN Changjun,YANG Zhao,WANG Chaozheng,LI Yixing,LI Fenfang,YUAN Debao,TAN Lin,CHOU Houyuan,CHEN Wenxue and JIN Zhiqiang.Prokaryotic Expression of Polyphenol Oxidase Mature Protein from Banana (Musa acuminate)[J].Chinese Journal of Tropical Crops,2015,36(12):2198-2203. |
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| Authors: | CHEN Jiao SUN Changjun YANG Zhao WANG Chaozheng LI Yixing LI Fenfang YUAN Debao TAN Lin CHOU Houyuan CHEN Wenxue and JIN Zhiqiang |
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| Institution: | Haikou Experimental Station, Chinese Academy of Tropical Agricultural Sciences/Hainan Key Laboratory of Banana Genetic Improvement;Haikou Experimental Station, Chinese Academy of Tropical Agricultural Sciences/Hainan Key Laboratory of Banana Genetic Improvement;Institute of Food, Hainan University;Haikou Experimental Station, Chinese Academy of Tropical Agricultural Sciences/Hainan Key Laboratory of Banana Genetic Improvement;Haikou Experimental Station, Chinese Academy of Tropical Agricultural Sciences/Hainan Key Laboratory of Banana Genetic Improvement;Haikou Experimental Station, Chinese Academy of Tropical Agricultural Sciences/Hainan Key Laboratory of Banana Genetic Improvement;Haikou Experimental Station, Chinese Academy of Tropical Agricultural Sciences/Hainan Key Laboratory of Banana Genetic Improvement;Haikou Experimental Station, Chinese Academy of Tropical Agricultural Sciences/Hainan Key Laboratory of Banana Genetic Improvement;Institute of Food, Hainan University;Institute of Food, Hainan University;Haikou Experimental Station, Chinese Academy of Tropical Agricultural Sciences/Hainan Key Laboratory of Banana Genetic Improvement |
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| Abstract: | A full-length PPO gene was isolated from banana, and the prokaryotic expression of this full-length gene was not succeeded. A 47 amino acids length transit peptide in its N-terminus was predicted by further amino acid sequence analysis. Prokaryotic expression of mature PPO protein without the transit peptide was performed. SDS-PAGE electrophoresis results showed a 62 ku specific protein bands with the expected size, and mass spectrometry analysis results showed that the recombinant protein was banana PPO. The results indicated that the transfer peptide may affect banana PPO expression in vitro, and lay some theory foundation to study the functions of banana PPO and its role in banana browning physiology and regulatory mechanisms. |
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