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PMID: 1701429 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S.

An expression vector for the archaebacterium Haloferax volcanii.

Journal of bacteriology ·Vol. 172 ·No. 12 ·1990-12-00 ·Pages 7104-10

Nieuwlandt DT, Daniels CJ

Abstract

The recent development of an efficient transformation method and shuttle vectors for Haloferax volcanii has set the stage for rapid progress in archaebacterial molecular biology. We describe a shuttle-expression vector that can be selected for and maintained in either H. volcanii or Escherichia coli and permits the expression of cloned genes in H. volcanii. The vector, pWL204, was constructed by incorporating an H. volcanii tRNA(Lys) gene promoter into a derivative of the H. volcanii-E. coli shuttle vector pWL102. The vector has been used to express a modified, intron-containing, H. mediterranei tRNA(Trp) gene (tRNA(Trp)-O167). Transcription from the tRNA(Lys) gene promoter in vivo was detected by Northern (RNA) analysis with an oligonucleotide probe complementary to the unique intron sequence of tRNA(Trp)-O167. Dependence of transcription on the tRNA(Lys) promoter was demonstrated by the absence of transcription when the promoter sequence was deleted from the vector and by mapping the transcription initiation site by primer extension.

MeSH Terms
Archaea/genetics Base Sequence Blotting, Northern Cloning, Molecular/methods DNA, Bacterial/genetics Escherichia coli/genetics Gene Expression Genetic Vectors Molecular Sequence Data Promoter Regions, Genetic RNA, Bacterial/genetics RNA, Transfer, Lys/genetics Restriction Mapping Transcription, Genetic Transformation, Genetic
Chemicals
DNA, Bacterial RNA, Bacterial RNA, Transfer, Lys
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Nieuwlandt D T
Department of Microbiology, Ohio State University, Columbus 43210.
Daniels C J
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31 references, click to expand
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.172.12.7104-7110.1990
Published
1990-12-00
Pages
7104-10
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC210833
Subset
IM
Databases
GENBANK
M62714, M62715
External Links
PubMed source DOI: 10.1128/jb.172.12.7104-7110.1990 PMC Full Text View journal details
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