[从脱嘧啶鲱鱼精DNA水解产物中制备分离单、二和三嘌呤核苷酸]

[Preparative isolation of mono-, di- and tripurine nucleotides from hydrolysates of depyrimidinated herring sperm DNA].

作者信息

Schott H, Schrade H

出版信息

J Chromatogr. 1983 Aug 19;265(2):257-75. doi: 10.1016/s0021-9673(01)96721-4.

Abstract

The purine nucleotides pdAp, pdGp, (dA)2, (dA-dG), (dG-dA), (dG)2, (dA)3, (dA-dG-dA), (dA-dA-dG), (dG-dA-dA), (dG-dA-dG) and known mixtures of purine nucleotide sequence isomers were separated by preparative scale chromatography of partial hydrolysates of depyrimidinated herring sperm DNA. Herring sperm DNA is first partially hydrolysed to a mixture of purine nucleotides. The low-molecular-weight oligonucleotides are then separated by column chromatography on DEAE-cellulose at pH 7.5, and fractionated by chromatography on QAE-Sephadex. Impurities which are not fully removed by column chromatography are separated by paper chromatography. The sequence of the isolated DNA fragments and the composition of the mixtures of sequence isomers were determined from the chromatographic data, absorption characteristics and by enzymatic degradation.

摘要

通过对脱嘧啶鲱鱼精DNA部分水解产物进行制备规模色谱法，分离出嘌呤核苷酸pdAp、pdGp、(dA)2、(dA - dG)、(dG - dA)、(dG)2、(dA)3、(dA - dG - dA)、(dA - dA - dG)、(dG - dA - dA)、(dG - dA - dG)以及已知的嘌呤核苷酸序列异构体混合物。鲱鱼精DNA首先被部分水解为嘌呤核苷酸混合物。然后，通过在pH 7.5的DEAE - 纤维素柱色谱分离低分子量寡核苷酸，并通过QAE - 葡聚糖凝胶柱色谱进行分级分离。未通过柱色谱完全去除的杂质通过纸色谱法分离。根据色谱数据、吸收特性以及酶促降解确定分离出的DNA片段的序列和序列异构体混合物的组成。