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。
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.
出处
《激光生物学报》
CAS
CSCD
1998年第2期150-153,共4页
Acta Laser Biology Sinica
基金
国家自然科学基金
关键词
脑钠肽
核磁共振波谱
样品制备
分离
纯化
brain natriuretic peptide(BNP) nuclear magnetic resonance(NMR) samples preparation separation purification.