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| 重组别藻蓝蛋白(rAPC)的纯化与鉴定 | |
| 引用本文： | 唐志红,李富超,吴少杰,葛保胜,林 凡,任育红,秦 松.重组别藻蓝蛋白(rAPC)的纯化与鉴定[J].海洋科学,2006,30(3):56-59. |
| 作者姓名： | 唐志红 李富超 吴少杰 葛保胜 林 凡 任育红 秦 松 |
| 作者单位： | 1. 中国科学院,海洋研究所,山东,青岛,266071;中国科学院,研究生院,北京,100039;烟台大学,海洋学院,山东,烟台,264005 2. 中国科学院,海洋研究所,山东,青岛,266071;中国科学院,研究生院,北京,100039 3. 中国科学院,海洋研究所,山东,青岛,266071 |
| 基金项目： | 国家863计划资助项目(2001AA6204130)，中国科学院知识创新项目(KZCX3–SW–215) |
| 摘 要： | 报道了重组大肠杆菌表达的别藻蓝蛋白的下游生产工艺研究,并对产物进行了分析鉴定。工程菌株P1先在NBSBioFlo3000型5L自动发酵罐进行高密度发酵,融合蛋白获得了高效表达,且主要在细胞内以可溶形式存在。纯化前先将获得的菌体超声破碎,然后经硫酸铵沉淀、疏水层析和离子交换层析三步纯化,接着对纯化的重组别藻蓝蛋白(rAPC)进行SDS-PAGE、PAGE、Westernblot等电点分析。证明已获得纯度为96.4%的rAPC,其等电点为6.0,并且具有与天然APC相似的抗原性。 |
| 关 键 词： | 工程菌培养 重组别藻蓝蛋白(rAPC) 纯化 |
| 文章编号： | 1000-3096（2006）03-0056-04 |
| 收稿时间： | 2004-04-25 |
| 修稿时间： | 2004-08-20 |
Purification and characterization of recombinant allophycoc-yanin(rAPC) expressed in a recombinant Escherichia coli P1 |
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| TANG Zhi-hong,LI Fu-chao,WU Shao-jie,GE Bao-sheng,LIN Fan,REN Yu-hong,QIN Song.Purification and characterization of recombinant allophycoc-yanin(rAPC) expressed in a recombinant Escherichia coli P1[J].Marine Sciences,2006,30(3):56-59. | |
| Authors: | TANG Zhi-hong LI Fu-chao WU Shao-jie GE Bao-sheng LIN Fan REN Yu-hong QIN Song |
| Institution: | 1 .Institute of Oceanology, the Chinese Academy of Sciences, Qingdao 266071 ,China;2. Graduate School, the Chinese Academy of Sciences, Beijing 100039,China;3. Ocean School, Yantai University, Yantai 264005,China |
| Abstract: | This study reports a method to purify rAPC and analysis of the purified rAPC.The culture of the recombinant Escherichia.coli strain was performed in a NBS BioFlo 3000 5 L fermentor(New Brunswick,USA);The final cell density was 16.8g dried cell/L broth and rAPC was produced as a water-soluble type.After sonication,the supernatant of free cell was fractionally precipited by ammonium sulfate.The protein was further purified by hydrophobic interaction chromatography and ion exchange chromatography.The purity of rAPC reached 96.4%;The purified rAPC was an acid protein(PI=6.0) and had a Mr value of approximate 60 ku;And the immunogenicity of rAPC is identical with that of its natural counterpart. |
| Keywords: | recombinant Allophycocyanin(rAPC) purification |
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