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PMID: 2996886 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

T4 polynucleotide kinase; cloning of the gene (pseT) and amplification of its product.

The EMBO journal ·Vol. 4 ·No. 10 ·1985-10-00 ·Pages 2695-703

Midgley CA, Murray NE

Abstract

The T4 gene (pseT) for polynucleotide kinase (pnk) has been cloned in lambda. Induction of a lambda E-W-S-cI857 prophage in which the pseT gene can be transcribed from the late lambda promoter, PR1, leads to greater than 100-fold amplification of pnk activity; pnk comprises approximately 7% of the total soluble cell protein. The purified enzyme, as expected, is both a 5'-kinase and a 3'-phosphatase. The amino acid sequence deduced from an open reading frame identified as the pseT gene contains a sequence which corresponds particularly well with that part of the adenine nucleotide binding site of adenylate kinase shown to form a flexible loop. A deletion mutant that lacks 5'-kinase activity, and possibly also 3'-phosphatase activity, has lost two amino acids from within the proposed loop structure. A second region of the pnk sequence shares homology with phosphoglycerate kinase, yeast inorganic pyrophosphatase and histone 2b from various organisms.

MeSH Terms
Amino Acid Sequence Base Sequence Cloning, Molecular Gene Amplification Gene Expression Regulation Genes Genes, Viral Molecular Weight Phosphotransferases/genetics Polynucleotide 5'-Hydroxyl-Kinase/genetics T-Phages/genetics Transcription, Genetic Viral Proteins/genetics Virus Replication
Chemicals
Viral Proteins Phosphotransferases Polynucleotide 5'-Hydroxyl-Kinase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Midgley C A
Murray N E
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47 references, click to expand
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1985-10-00
Pages
2695-703
Language
English
Region
England
NLM ID
8208664
PMCID
PMC554562
Subset
IM
Databases
GENBANK
X03007
External Links
PubMed source PMC Full Text View journal details
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