| 欧李ChPSY基因的原核表达 |
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| 引用本文： | 陈俊奇,王鹏飞,杜俊杰,李润丰,洪文泓,李佩,马精选,张建成.欧李ChPSY基因的原核表达[J].山西农业科学,2012,40(12). |
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| 作者姓名： | 陈俊奇 王鹏飞 杜俊杰 李润丰 洪文泓 李佩 马精选 张建成 |
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| 作者单位： | 山西农业大学园艺学院,山西 太谷,030801 |
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| 基金项目： | 高等学校博士学科点专项科研基金，山西省自然科学基金项目，山西省人力资源和社会保障厅优秀人才专项补助基金，山西省高等学校大学生创新创业训练项目，山西农业大学大学生科技创新项目 |
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| 摘 要： | 为了解欧李果实类胡萝卜素合成过程中八氢番茄红素合成酶基因的作用和功能,将从欧李成熟果实中分离的ChPSY cDNA序列,连接到原核表达载体pET-28a(+),导入大肠杆菌BL21(DE3)体内诱导表达。SDS-PAGE分析表明,1.0 mmol/LIPTG诱导4 h,表达了相对分子量约为45.8 kD融合蛋白产物;IPTG的浓度和诱导时间优化表明,1.2 mmol/LIPTG诱导6 h时,融合蛋白的表达量最大。此外,Western-blot试验证实,表达的融合蛋白能与抗6×His的单抗发生特异性反应。这为欧李八氢番茄红素合成酶基因ChPSY生物学功能的深入研究奠定了基础。
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| 关 键 词： | 欧李 八氢番茄红素合成酶 cDNA序列 原核表达 |
Prokaryotic Expression of ChPSY Gene from Cerasus humilis |
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| CHEN Jun-qi
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WANG Peng-fei
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DU Jun-jie
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LI Run-feng
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HONG Wen-hong
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LI Pei
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MA Jing-xuan
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ZHANG Jian-cheng.Prokaryotic Expression of ChPSY Gene from Cerasus humilis[J].Journal of Shanxi Agricultural Sciences,2012,40(12). |
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| Authors: | CHEN Jun-qi WANG Peng-fei DU Jun-jie LI Run-feng HONG Wen-hong LI Pei MA Jing-xuan ZHANG Jian-cheng |
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| Abstract: | To understand the role and function of the Cerasus humilis phytoene synthase gene(ChPSY),the full-length coding region of ChPSY cDNA from the Cerasus humilis mature fruit was linked into the prokaryotic expression vector pET-28a(+).The recombinant plasmid pET-ChPSY was transformed into E.coli BL21(DE3) for induced expression.The analysis of SDS-PAGE demonstrated that the fusion protein(6×His-PSY) was produced at 4h after induction by 1.0 mmol/L IPTG and migrated at a size of about 45.8 kD.The optimization of IPTG concentration and induction time showed the fusion protein quantity was maximal after 6 h induction by 1.2 mmol/L IPTG.The results of Western-blot demonstrated that the fusion protein(6×His-PSY) could be recognized by anti-6×His monoclonal antibody.The research provided a basis for further study on the biological function of ChPSY gene in fruit carotenoid biosynthesis of Cerasus humilis(Bge) Sok. |
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| Keywords: | Cerasus humilis phytoene synthase cDNA sequence prokaryotic expression |
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