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核磁共振波谱研究脑钠肽空间结构样品制备

The Samples of BNP was been Maked for Studing the Structure of BNP with NMR

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【作者】 傅蕙英曾王勇刘竞

【Author】 Fu Huiying* Zen Wangyong Liu Jin(University of science and Technology of china,Hefie 230026)

【机构】 中国科学技术大学生物系

【摘要】 用多维核磁共振波谱(NMR)研究脑钠肽(BNP)空间结构时,必需制备大量的BNP样品。用pGEX—3X作载体与BNP—38cDNA构建成能高效表达GST—NP—38融合蛋白的工程菌,表达出的融合蛋白用珠(GST—agarosebeads)分离,动态分离的效果是静态分离的三倍;分离时,由振荡2分钟延至30分钟,分离量可增加14倍。纯化GST—BNP—38时,动态洗脱的得率是静态洗脱的5.2倍,在动态体系纯化的得率是在静态体系的10倍。因此,分离纯化GST—BNP—38时,用珠振荡吸附30分钟,再用GSH振荡洗脱30分钟,可使其得率提高60倍以上。这为NMR研究BNP结构时大量样品制备奠定了重要基础。

【Abstract】 When the structure of BNP is been studing by NMR,the BNP samples must be made in larg amount.The engineering bacterium has been constructed with the pGEX-3x as a vector and the BNP-38 cDNA as a foreign DNA that expresses the GST-BNP-38 fusion protien.The fusion protein can be separated with GST-agarose beads.The yeild of separation of fusion protein umder rotation condition is 3 times that under non-rotation condition.When the rotating time was increadsed to 30′,the separating qnentity has been increased by 14 times as against that the time was 2′,When the GST -BNP -38 were purificated ,the eluting amount under rotaion condition 52 times that under non-rotation condition.The purification yeild of fusion protein in a rotating system is 10 times that in a non-rotating system.As a result ,when fusion proteins were absorbed for 30′again under rotation condition,the yeild of purification fusion proteins may be increased by 60 times as against that under non-rotation condition.This has set up an important foundation that preparate samples in larg amount for the research of BNP with NMR.

【基金】 国家自然科学基金
  • 【文献出处】 激光生物学报 ,ACTA LASER BIOLOGY SINICA , 编辑部邮箱 ,1998年02期
  • 【分类号】R338,R445.2
  • 【被引频次】4
  • 【下载频次】61
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