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Formation of an RNA primer for initiation of replication of ColE1 DNA by ribonuclease H.
Proc Natl Acad Sci U S A. 1980 May; 77(5):2450-4.PN

Abstract

A plasmid that consists of an 812-base-pair segment containing the replication origin of plasmid ColE1 and of a 1240-base-pair segment containing a beta-lactamase gene has been constructed. The plasmid DNA has three principal sites where transcription is initiated in vitro. One is located in the ColE1 segment 555 nucleotides upstream from the origin. Most transcription from this site extends past the origin; some of the transcripts form hybrids spontaneously with the template at their 3' portions. Cleavage of these transcripts by RNase H generates 3' termini at the origin region. When DNA polymerase I is included in the reaction along with RNA polymerase and RNase H, dAMP or dCMP is added directly onto the cleaved RNA molecules, most of which retain the intact 5' terminus. The addition of a deoxyribonucleotide to the cleaved RNA can be regarded as the first step of ColE1 DNA synthesis. Once it has served as a primer, the RNA is eliminated from the product by RNase H.

Authors

No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

6156450

Citation

Itoh, T, and J Tomizawa. "Formation of an RNA Primer for Initiation of Replication of ColE1 DNA By Ribonuclease H." Proceedings of the National Academy of Sciences of the United States of America, vol. 77, no. 5, 1980, pp. 2450-4.
Itoh T, Tomizawa J. Formation of an RNA primer for initiation of replication of ColE1 DNA by ribonuclease H. Proc Natl Acad Sci USA. 1980;77(5):2450-4.
Itoh, T., & Tomizawa, J. (1980). Formation of an RNA primer for initiation of replication of ColE1 DNA by ribonuclease H. Proceedings of the National Academy of Sciences of the United States of America, 77(5), 2450-4.
Itoh T, Tomizawa J. Formation of an RNA Primer for Initiation of Replication of ColE1 DNA By Ribonuclease H. Proc Natl Acad Sci USA. 1980;77(5):2450-4. PubMed PMID: 6156450.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Formation of an RNA primer for initiation of replication of ColE1 DNA by ribonuclease H. AU - Itoh,T, AU - Tomizawa,J, PY - 1980/5/1/pubmed PY - 1980/5/1/medline PY - 1980/5/1/entrez SP - 2450 EP - 4 JF - Proceedings of the National Academy of Sciences of the United States of America JO - Proc. Natl. Acad. Sci. U.S.A. VL - 77 IS - 5 N2 - A plasmid that consists of an 812-base-pair segment containing the replication origin of plasmid ColE1 and of a 1240-base-pair segment containing a beta-lactamase gene has been constructed. The plasmid DNA has three principal sites where transcription is initiated in vitro. One is located in the ColE1 segment 555 nucleotides upstream from the origin. Most transcription from this site extends past the origin; some of the transcripts form hybrids spontaneously with the template at their 3' portions. Cleavage of these transcripts by RNase H generates 3' termini at the origin region. When DNA polymerase I is included in the reaction along with RNA polymerase and RNase H, dAMP or dCMP is added directly onto the cleaved RNA molecules, most of which retain the intact 5' terminus. The addition of a deoxyribonucleotide to the cleaved RNA can be regarded as the first step of ColE1 DNA synthesis. Once it has served as a primer, the RNA is eliminated from the product by RNase H. SN - 0027-8424 UR - https://www.unboundmedicine.com/medline/citation/6156450/Formation_of_an_RNA_primer_for_initiation_of_replication_of_ColE1_DNA_by_ribonuclease_H_ L2 - https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/6156450/ DB - PRIME DP - Unbound Medicine ER -