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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
【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 52 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.
【Key words】 brain natriuretic peptide(BNP) nuclear magnetic resonance(NMR) samples preparation separation purification.;
- 【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