TERMINAL PROTEIN-PRIMED DNA AMPLIFICATION

被引：67

作者：

BLANCO, L ^{[1
]}

LAZARO, JM ^{[1
]}

DEVEGA, M ^{[1
]}

BONNIN, A ^{[1
]}

SALAS, M ^{[1
]}

机构：

[1] UNIV AUTONOMA MADRID,CSIC,CTR BIOL MOLEC SEVERO OCHOA,E-28049 MADRID,SPAIN

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 1994年 / 91卷 / 25期

关键词：

D O I：

10.1073/pnas.91.25.12198

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：

By using appropriate amounts of four bacteriophage phi 29 DNA replication proteins-terminal protein, DNA polymerase, protein p6 (double-stranded DNA-binding protein), and protein p5 (single-stranded DNA-binding protein)-it has been possible to amplify limited amounts of the 19,285-bp-long phi 29 DNA molecule by three orders of magnitude after 1 hr of incubation at 30 degrees C. Moreover, the quality of the amplified material was demonstrated by transfection experiments, in which infectivity of the synthetic (amplified) phi 29 DNA, measured as the ability to produce phage particles, was identical to that of the natural phi 29 DNA obtained from virions. The results presented in this paper establish some of the requisites for the development of isothermal DNA amplification strategies based on the bacteriophage phi 29 DNA replication machinery that are suitable for the amplification of very large (>70 kb) segments of DNA.

引用

页码：12198 / 12202

页数：5

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