| 鱼腥蓝细菌PCC 7120中两个磷酸酶N端结构域活性分析 |
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| 引用本文： | 许兴俭,陈雯莉,王莉. 鱼腥蓝细菌PCC 7120中两个磷酸酶N端结构域活性分析[J]. 湖北农业科学, 2012, 51(7): 1474-1477 |
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| 作者姓名： | 许兴俭 陈雯莉 王莉 |
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| 作者单位： | 华中农业大学生命科学技术学院/农业微生物学国家重点实验室,武汉,430070 |
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| 摘 要： | 为了研究鱼腥蓝细菌(A nabaena sp.)PCC 7120中两个PP2C类蛋白磷酸酶PrpJ1和PrpJ2的磷酸酶活性,将编码两个蛋白N端至跨膜区的基因片段重组到质粒中,转化大肠杆菌进行原核表达,获得了可溶性的PrpJ1up和PrpJ2up蛋白.并且以pNPP为底物测定了所得蛋白的磷酸酶活性,双倒数作图法结果显示PrpJ1up蛋白的KK为0.30 mmol/L,Vmax为7.10 nmol/min；PrpJ2up蛋白的Km为0.24 mmol/L,Vmax为0.43 nmol/min.
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| 关 键 词： | 鱼腥蓝细菌(Anabaena sp.) PrpJ1 PrpJ2 N端结构域 磷酸酶活性 |
Activity Determination of N-domain of Two Phosphatase from Anabaena sp.PCC 7120 |
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| XU Xing-jian , CHEN Wen-li , WANG Li. Activity Determination of N-domain of Two Phosphatase from Anabaena sp.PCC 7120[J]. Hubei Agricultural Sciences, 2012, 51(7): 1474-1477 |
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| Authors: | XU Xing-jian CHEN Wen-li WANG Li |
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| Affiliation: | (College of Life Science and Technology/State Key Laboratory of Agricultural Microbiology,Huazhong Agricultural University, Wuhan 430070,China) |
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| Abstract: | The gene fragments encoding N-domain of two phosphatase proteins PrpJ1 and PrpJ2 were cloned into pET28a.Soluble proteins PrpJ1up and PrpJ2up were obtained by prokaryotic expression,and then purified by Ni-NTA argrose.The kinetic constants of PrpJ1up and PrpJ2up towards pNPP were determined.The results showed that Km and Vmax of PrpJ1up were 0.30 mmol/L and 7.10 nmol/min respectively;While Km and Vmax of PrpJ2up were 0.24 mmol/L and 0.43 nmol/min respectively. |
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| Keywords: | Anabaena sp.PCC 7120 PrpJ1 PrpJ2 N-domain phosphatase activity |
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