RAPID PURIFICATION OF HIGHLY-ACTIVE RIBOSOMES FROM ESCHERICHIA-COLI
RAPID PURIFICATION OF HIGHLY-ACTIVE RIBOSOMES FROM ESCHERICHIA-COLI
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DOI：
10.1016/0003-2697(80)90472-8
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发表时间：
1980-01-01
影响因子：
2.9
通讯作者：
JELENC, PC
中科院分区：
文献类型：
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作者：
JELENC, PC
高活性盐洗核糖体来自E.通过在Sephacryl S-200上在含有1 M NH 4Cl和腐胺、亚精胺、Ca 2+和Mg 2+的缓冲液中进行凝胶过滤来分离大肠杆菌。可在不到24小时内处理多达数百克的细胞。在甲醇中制备的30%（体积/体积）的核糖体溶液最好作为液体储存在-20 ℃。C.本发明描述了一种改进的多聚（U）依赖性肽合成测定法，其中苯丙氨酸聚合大大增强，而亮氨酸的错误掺入保持在体内水平。
Highly active salt-washed ribosomes from E. coli are isolated by gel filtration on Sephacryl S-200 in a buffer containing 1 M NH4Cl and putrescine, spermidine, Ca2+ and Mg2+. Up to several hundred grams of cells can be processed in less than 24 h. The ribosome solution made 30% (vol/vol) in methanol is best stored as a liquid at -20.degree. C. An improved poly(U)-dependent peptide synthesis assay is described in which phenylalanine polymerization is greatly enhanced while the misincorporation of leucine is kept at in vivo levels.