NONHOMOLOGOUS DNA END JOINING OF SYNTHETIC HAIRPIN SUBSTRATES IN XENOPUS-LAEVIS EGG EXTRACTS

被引：20

作者：

BEYERT, N ^{[1
]}

REICHENBERGER, S ^{[1
]}

PETERS, M ^{[1
]}

HARTUNG, M ^{[1
]}

GOTTLICH, B ^{[1
]}

GOEDECKE, W ^{[1
]}

VIELMETTER, W ^{[1
]}

PFEIFFER, P ^{[1
]}

机构：

[1] UNIV COLOGNE,INST GENET,D-50674 COLOGNE,GERMANY

来源：

NUCLEIC ACIDS RESEARCH | 1994年 / 22卷 / 09期

关键词：

D O I：

10.1093/nar/22.9.1643

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Processes of DNA end joining are assumed to play a major role in the elimination of DNA double-strand breaks (DSB) in higher eucaryotic cells. Linear plasmid molecules terminated by nonhomologous restriction ends are the typical substrates used in the analysis of joining mechanisms. However, due to their limited structural variability, DSB ends generated by restriction cleavage cover probably only part of the total spectrum of naturally occurring DSB termini. We therefore devised novel DNA substrates consisting of synthetic hairpin-shaped oligonucleotides which permit the construction of blunt ends and 5'- or 3'-protruding single-strands (PSS) of arbitrary sequence and length. These substrates were tested in extracts of Xenopus laevis eggs known to efficiently join linear plasmids bearing nonhomologous restriction termini (Pfeiffer and Vielmetter, 1988). Sequences of hairpin junctions indicate that the short hairpins are joined by the same mechanisms as the plasmid substrates. However, the bimolecular DNA end joining reaction was only detectable when both hairpin partners had a minimal duplex stem length of 27bp and their PSS-tails did not exceed 10nt.

引用

页码：1643 / 1650

页数：8

共 34 条

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