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PMID: 6272270 Published · ppublish English Journal Article

Molecular cloning and amplification of the adenylate cyclase gene.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 78 ·No. 8 ·1981-08-00 ·Pages 4684-8

Wang JY, Clegg DO, Koshland DE

Abstract

A segment of DNA containing cya, the gene for adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1], has been isolated from Salmonella typhimurium. The phage lambda gt4 was used as a cloning vector and adenylate cyclase-positive hybrid phages were isolated that complemented adenylate cyclase-negative bacteria. The cloned DNA fragment encodes a polypeptide of molecular weight 81,000 that gives rise to adenylate cyclase activity. This protein represents a functional mutant of the bacterial adenylate cyclase. When the cya gene was amplified by inserting into a multicopy plasmid, the enzyme activity was overproduced 20-fold, but the cyclic AMP level increased only 60%, suggesting several probable regulatory mechanisms. Overproduction of enzymes by recombinant DNA techniques can be a useful probe of relationships in the metabolizing organism in vivo.

MeSH Terms
Adenylyl Cyclases/genetics Bacteriophage lambda/genetics Chromosome Mapping Cloning, Molecular/methods Cyclic AMP/metabolism Gene Amplification Gene Expression Regulation Genes Molecular Weight Operon Plasmids Salmonella typhimurium/genetics
Chemicals
Cyclic AMP Adenylyl Cyclases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Wang J Y
Clegg D O
Koshland D E
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34 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.78.8.4684
Published
1981-08-00
Pages
4684-8
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC320227
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.78.8.4684 PMC Full Text View journal details
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