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PMID: 3511029 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Cloning, overproduction, and purification of the B2 subunit of ribonucleoside-diphosphate reductase.

Journal of bacteriology ·Vol. 165 ·No. 2 ·1986-02-00 ·Pages 363-6

Salowe SP, Stubbe J

Abstract

The nrdB gene, which encodes the B2 subunit of Escherichia coli ribonucleotide reductase (EC 1.17.4.1), was cloned into multicopy plasmid pSPS2. This vector, which contains the pL promoter of bacteriophage lambda and the tetracycline resistance gene of pBR322, was transformed into a lysogenic host with a thermolabile repressor. In the newly constructed strain, subunit B2 constituted approximately 25% of the soluble protein after heat induction, an overproduction of several hundredfold relative to the wild-type strain. Purification to homogeneity of the overproduced protein was accomplished by using DEAE and quaternary aminoethyl ion-exchange resins.

MeSH Terms
Amino Acid Sequence Cloning, Molecular Escherichia coli/enzymology Gene Expression Regulation Macromolecular Substances Ribonucleoside Diphosphate Reductase/biosynthesis,genetics,isolation & purification Ribonucleotide Reductases/genetics
Chemicals
Macromolecular Substances Ribonucleotide Reductases Ribonucleoside Diphosphate Reductase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Salowe S P
Stubbe J
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23 references, click to expand
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.165.2.363-366.1986
Published
1986-02-00
Pages
363-6
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC214425
Subset
IM
Grants
NIADDK NIH HHS · AM 01222 · United States
NIGMS NIH HHS · GM 29595 · United States
External Links
PubMed source DOI: 10.1128/jb.165.2.363-366.1986 PMC Full Text View journal details
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