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PMID: 2429304 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

A temperature-dependent pBR322 copy number mutant resulting from a Tn5 position effect.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 83 ·No. 19 ·1986-10-00 ·Pages 7381-5

Lupski JR, Projan SJ, Ozaki LS, Godson GN

Abstract

In the process of randomly mutagenizing a recombinant pBR322 clone with transposon Tn5, a high copy number plasmid mutant, pLO88, has been isolated. The copy number phenotype of pLO88 is observed only at elevated temperatures, greater than or equal to 37 degrees C, and is due to the precise position of a Tn5 insertion. Nucleotide sequence of the Tn5-pBR322 junction reveals that Tn5-88 has inserted into an open reading frame that codes for a 63 amino acid protein previously shown to negatively regulate pBR322 plasmid copy number. By deleting portions of the Tn5 it is shown that the copy number phenotype is due not only to the insertion of Tn5 in pBR322 but also to the requirement that some Tn5 sequences remain intact. It appears that an outwardly directed Tn5 promoter initiates the synthesis of a transcript (RNA X) that interferes with the normal repressor RNA (RNA I)-primer RNA (RNA II) interaction at elevated temperatures.

MeSH Terms
DNA Replication DNA Transposable Elements Mutation Plasmids RNA, Bacterial/genetics Temperature
Chemicals
DNA Transposable Elements RNA, Bacterial
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Lupski J R
Projan S J
Ozaki L S
Godson G N
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.83.19.7381
Published
1986-10-00
Pages
7381-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC386721
Subset
IM
Grants
PHS HHS · 7-1142-996 · United States
NIGMS NIH HHS · GM33278 · United States
External Links
PubMed source DOI: 10.1073/pnas.83.19.7381 PMC Full Text View journal details
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