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大肠杆菌DNA聚合酶I介导的逆转录

Reverse transcription by Escherichia coli DNA polymerase I.

作者信息

Karkas J D

出版信息

Proc Natl Acad Sci U S A. 1973 Dec;70(12):3834-8. doi: 10.1073/pnas.70.12.3834.

Abstract

E. coli DNA polymerase I (EC 2.7.7.7) can engage in either DNA- or RNA-directed DNA synthesis with hybrid templates. The choice of the strand to be transcribed depends primarily on the relative lengths of the two strands of the hybrid, the longer strand serving as the template and the shorter as the primer. If a polynucleotide is reduced in size by exposure to an endonuclease before being hybridized to the complementary strand, the template efficiency of the latter increases several-fold. Under properly selected conditions, highly efficient reverse transcription of the all-ribonucleotide template-primers poly(A).oligo(U), poly(C).oligo(I), and poly(I).oligo(C) can be achieved. "f1 RNA," the RNA strand of an f1 DNA.RNA hybrid, can also serve as template for reverse transcription either after "nicking" of the hybrid with DNase, or after separation from the DNA strand and priming by DNase-treated f1 DNA.

摘要

大肠杆菌DNA聚合酶I(EC 2.7.7.7)可以利用杂交模板进行DNA或RNA指导的DNA合成。转录链的选择主要取决于杂交双链中两条链的相对长度,较长的链作为模板,较短的链作为引物。如果一个多核苷酸在与互补链杂交之前通过核酸内切酶处理而减小尺寸,那么后者的模板效率会提高几倍。在适当选择的条件下,可以实现全核糖核苷酸模板引物聚(A)·寡聚(U)、聚(C)·寡聚(I)和聚(I)·寡聚(C)的高效逆转录。f1 DNA·RNA杂交体的RNA链“f1 RNA”,在杂交体被DNase“切口”后,或与DNA链分离并用DNase处理的f1 DNA引发后,也可以作为逆转录的模板。

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Reverse transcription by Escherichia coli DNA polymerase I.大肠杆菌DNA聚合酶I介导的逆转录
Proc Natl Acad Sci U S A. 1973 Dec;70(12):3834-8. doi: 10.1073/pnas.70.12.3834.

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