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[合成寡脱氧核苷酸在限制性内切酶MspI研究中的应用]

[Synthetic oligodeoxynucleotides in the study of restriction endonuclease MspI].

作者信息

Boreskov Iu G, Titeeva G R, Berlin Iu A

出版信息

Bioorg Khim. 1988 Mar;14(3):333-9.

PMID:2454631
Abstract

Interaction of MspI restriction endonuclease with a series of oligodeoxynucleotides, varying in stability of secondary structure and in location of the restriction site, has been studied. It is shown that a functionally active MspI-site must be double-stranded and flanked from both sides. Separate MspI-cleavage of dodecanucleotides dCGACCCGGGATC and dGATCCCGGGTCG is inhibited by the reaction products as well as by non-homological hexanucleotides dGGTACC and dGGATCC (but not by dCGGCGC). Polyethylene glycol in low concentrations (1-3%) promotes and in higher concentrations (7-14%) inhibits the cleavage. A scheme of MspI functioning is suggested including enzyme's step-by-step recognition of the restriction site and its nonspecific interaction with flanking segments of DNA, which leads to formation of the productive complex.

摘要

已研究了MspI限制性内切酶与一系列寡脱氧核苷酸的相互作用,这些寡脱氧核苷酸在二级结构稳定性和限制位点位置上有所不同。结果表明,功能活性的MspI位点必须是双链的,且两侧都有侧翼。十二聚体dCGACCCGGGATC和dGATCCCGGGTCG的单独MspI切割受到反应产物以及非同源六聚体dGGTACC和dGGATCC(但不受dCGGCGC)的抑制。低浓度(1-3%)的聚乙二醇促进切割,而高浓度(7-14%)则抑制切割。提出了MspI的作用机制,包括酶对限制位点的逐步识别及其与DNA侧翼片段的非特异性相互作用,这导致形成有活性的复合物。

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