从面包酵母中提取和纯化半胱氨酸转移核糖核酸。
The extraction and purification of a cysteine transfer ribonucleic acid from baker's yeast.
作者信息
Holness N J, Atfield G
出版信息
Biochem J. 1976 Feb 1;153(2):429-35. doi: 10.1042/bj1530429.
Abstract
- A modification of the RPC 1 system of A.D. Kelmers, G.D. Novelli & M.P. Stulberg (1965) (J. Biol. Chem. 240, 3979-3983) is described in which the support medium is a Celite of narrow range particle size treated with dichlorodimethylsilane. 2. By using this system an apparently pure preparation of tRNA Cys was isolated from baker's yeast tRNA. 3. This preparation accepted at least 60% of the theoretical quantity of [3-14C]cysteine in a conventional assay and failed to accept isoleucine, phenylalanine, proline, serine or tyrosine. 4. A theoretical countercurrent-distribution curve calculated by assuming a distribution coefficient K of 2.03 was in excellent agreement with the profiles of E260 and cysteine-acceptor ability after 537 transfers in the 1.85 M-phosphate/formamide/propan-2-ol system of C.M. Connelly & B.P. Doctor (1965) (J. Biol. Chem. 241, 715-719). 5. Chromatography of tRNA Cys on Bio-Gel P100 polyacrylamide beads afforded two components one of which was far less efficient than the other in accepting cysteine. The base compositions of the two were similar.
摘要
- 描述了对A.D. 凯尔默斯、G.D. 诺维利和M.P. 斯图尔伯格（1965年）的RPC 1系统（《生物化学杂志》240, 3979 - 3983）的一种改进，其中支持介质是用二氯二甲基硅烷处理过的窄粒度范围的硅藻土。2. 通过使用该系统，从面包酵母tRNA中分离出了一种看似纯的tRNA Cys制剂。3. 在常规测定中，该制剂接受了至少60%理论量的[3 - 14C]半胱氨酸，且不接受异亮氨酸、苯丙氨酸、脯氨酸、丝氨酸或酪氨酸。4. 通过假设分配系数K为2.03计算出的理论逆流分布曲线，与在C.M. 康奈利和B.P. 多克特（1965年）（《生物化学杂志》241, 715 - 719）的1.85 M - 磷酸盐/甲酰胺/丙 - 2 - 醇系统中进行537次转移后的E260和半胱氨酸接受能力曲线非常吻合。5. tRNA Cys在Bio - Gel P100聚丙烯酰胺珠上进行色谱分析得到两个组分，其中一个在接受半胱氨酸方面比另一个效率低得多。两者的碱基组成相似。
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引用本文的文献
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The nucleotide sequence of cysteine transfer ribonucleic acid from baker's yeast. Identification of the products from partial degradation of the molecule and derivation of the complete sequence.面包酵母半胱氨酸转移核糖核酸的核苷酸序列。该分子部分降解产物的鉴定及完整序列的推导。
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