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PMID: 3534890 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

beta-Glucuronidase from Escherichia coli as a gene-fusion marker.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 83 ·No. 22 ·1986-11-00 ·Pages 8447-51

Jefferson RA, Burgess SM, Hirsh D

Abstract

We have developed a gene-fusion system based on the Escherichia coli beta-glucuronidase gene (uidA). The uidA gene has been cloned from E. coli K-12 and its entire nucleotide sequence has been determined. beta-Glucuronidase has been purified to homogeneity and characterized. The enzyme has a subunit molecular weight of 68,200, is very stable, and is easily and sensitively assayed using commercially available substrates. We have constructed gene fusions of the E. coli lacZ promoter and coding region with the coding region of the uidA gene that show beta-glucuronidase activity under lac control. Plasmid vectors have been constructed to facilitate the transfer of the beta-glucuronidase coding region to heterologous control regions, using many different restriction endonuclease cleavage sites. There are several biological systems in which uidA-encoded beta-glucuronidase may be an attractive alternative or complement to previously described gene-fusion markers such as beta-galactosidase or chloramphenicol acetyltransferase.

MeSH Terms
Base Sequence Chromosome Mapping Cloning, Molecular Escherichia coli/enzymology Genetic Engineering Genetic Markers Genetic Vectors Glucuronidase/genetics,isolation & purification
Chemicals
Genetic Markers Glucuronidase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Jefferson R A
Burgess S M
Hirsh D
References (20)
20 references, click to expand
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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.83.22.8447
Published
1986-11-00
Pages
8447-51
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC386947
Subset
IM
Grants
NIGMS NIH HHS · GM26515 · United States
Databases
GENBANK
M14641
External Links
PubMed source DOI: 10.1073/pnas.83.22.8447 PMC Full Text View journal details
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