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PMID: 6227808 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Isolation and preliminary characterization of the Chinese hamster thymidine kinase gene.

Molecular and cellular biology ·Vol. 3 ·No. 10 ·1983-10-00 ·Pages 1815-23

Lewis JA, Shimizu K, Zipser D

Abstract

The Chinese hamster thymidine kinase (TK) gene has been isolated from a recombinant phage library constructed with genomic DNA from mouse Ltk- cells transformed to Tk+ by transfection with Chinese hamster genomic DNA. The phage library was screened by the Benton-Davis plaque hybridization technique, using as probes, subclones of recombinant phage that were isolated from mouse Ltk+ transformants by the tRNA suppressor rescue method. The Chinese hamster TK gene is contained within 13.2 kilobases of genomic DNA in the isolate designated lambda 34S4. This gene, defined by restriction enzyme sensitivity experiments, homology studies with the chicken TK gene, and mRNA blotting experiments, may extend over 8.5 kilobases. Subclones of the lambda 34S4 isolate used as hybridization probes identified a 1,400-nucleotide polyadenylated RNA as the hamster TK mRNA. The abundance of this mRNA varies dramatically in Chinese hamster cells cultured under various growth conditions, providing direct evidence that the growth dependence of TK activity may be regulated in an important way at the level of cytoplasmic TK mRNA.

MeSH Terms
Animals Bacteriophage lambda/genetics Cloning, Molecular Cricetinae Cricetulus DNA, Recombinant/isolation & purification Female Genes L Cells/analysis Mice RNA, Messenger/genetics Thymidine Kinase/genetics
Chemicals
DNA, Recombinant RNA, Messenger Thymidine Kinase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Lewis J A
Shimizu K
Zipser D
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42 references, click to expand
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.3.10.1815-1823.1983
Published
1983-10-00
Pages
1815-23
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC370043
Subset
IM
Grants
NCI NIH HHS · 5POICA29569 · United States
NCI NIH HHS · CA09311 · United States
External Links
PubMed source DOI: 10.1128/mcb.3.10.1815-1823.1983 PMC Full Text View journal details
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