STRUCTURE-SPECIFIC ENDONUCLEOLYTIC CLEAVAGE OF NUCLEIC-ACIDS BY EUBACTERIAL DNA-POLYMERASES

被引:305
作者
LYAMICHEV, V [1 ]
BROW, MAD [1 ]
DAHLBERG, JE [1 ]
机构
[1] UNIV WISCONSIN,SCH MED,DEPT BIOMOLEC CHEM,1300 UNIV AVE,MADISON,WI 53706
关键词
D O I
10.1126/science.7683443
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Previously known 5' exonucleases of several eubacterial DNA polymerases have now been shown to be structure-specific endonucleases that cleave single-stranded DNA or RNA at the bifurcated end of a base-paired duplex. Cleavage was not coupled to synthesis, although primers accelerated the rate of cleavage considerably. The enzyme appeared to gain access to the cleavage site by moving from the free end of a 5' extension to the bifurcation of the duplex, where cleavage took place. Single-stranded 5' arms up to 200 nucleotides long were cleaved from such a duplex. Essentially any linear single-stranded nucleic acid can be targeted for specific cleavage by the 5' nuclease of DNA polymerase through hybridization with an oligonucleotide that converts the desired cleavage site into a substrate.
引用
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页码:778 / 783
页数:6
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