| 平颏海蛇PLA2在大肠杆菌中的融合表达 |
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| 引用本文： | 杨文利,叶兰汀,邹兰,汪枫,徐安龙.平颏海蛇PLA2在大肠杆菌中的融合表达[J].微生物学报,2002,42(4):400-405. |
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| 作者姓名： | 杨文利 叶兰汀 邹兰 汪枫 徐安龙 |
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| 作者单位： | 1. 中山大学生命科学学院生化系,海洋生物功能基因组开放实验室,广州,510275
2. 中国科技大学生命科学院,合肥,230027 |
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| 基金项目： | 国家高新技术海洋 86 3项目 ( 2 0 0 1AA6 2 6 0 1 0 )，国家自然科学基金重点项目 ( 6 9935 0 2 0 )~~ |
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| 摘 要： | 将编码平颏海蛇磷脂酶A2的基因(PLA29)分别克隆于硫氧环蛋白基因融合表达载体pThioHisC和pTRX的HP\|trxA和trxA基因的3′末端，构建符合读码框的融合表达载体pThioHisC\|PLA2和pTRX\|PLA2。25℃下经IPTG诱导，PLA2融合蛋白在两种原核系统中均能获得较高表达，但PLA2在pTRX系统下的表达量和蛋白溶解性优于pThioHisC系统。将两种系统下的表达产物进行金属螯合亲和层析纯化，Trx\|PLA2融合蛋白的纯度可达85%以上，而HPTrxPLA2不表现出对介质的亲和性，难以得到纯化。由此，将pTRX\|PLA2载体系统确立为进一步大量表达和纯化的载体系统。
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| 关 键 词： | 平颏海蛇磷脂酶A2， 融合表达， 亲和层析 |
| 文章编号： | 0001-6209(2002)04-0400-06 |
Fusion Expression of PLA -2 Gene From Lapemis hardwickii in E.coli |
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| Yang Wenli\\ Ye Lanting\\ Zou Lan\\ Wang Feng\\ Xu Anlong.Fusion Expression of PLA -2 Gene From Lapemis hardwickii in E.coli[J].Acta Microbiologica Sinica,2002,42(4):400-405. |
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| Authors: | Yang Wenli\\ Ye Lanting\\ Zou Lan\\ Wang Feng\\ Xu Anlong |
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| Institution: | National Opening Laboratory for Marine Functional Genomics Research, School of Life Science, Zhongshan University, Guangzhou 510275, China. |
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| Abstract: | The gene encoding PLA2(PLA2-9) from Lapemis hardwickii Gray venom was cloned to the 3' and of the thioredoxin gene (HP-trxA and trxA) in plasmid pthioHisC and pTRX to construct the pThioHisC-PLA2, and pTRX-PLA2 fusion expression vector. The fusion protein of PLA2 can be expressed in the two different systems induced by IPTG at 25 degrees C, but the expression level and the solubility of the fusion protein in pTRX were better than that in pThioHisC. The expressed product in the two systems were purified by immobilized metal-chelate affinity chromatography. The Trx-PLA2 fusion protein with over 85% purity was obtained and HP-Trx-PLA2 fusion protein can not be purified since it dose not exhibit affinity to the medium. So, the pTRX-PLA2 vector system was established for the large-scale expression and purification. |
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| Keywords: | PLA\-2 from Lapemis hardwickii Fusion expression Affinity chromatography |
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