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Cloning and Expression of the Adenosine Kinase Gene from Rat and Human Tissues,☆☆

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Abstract

Adenosine kinase is ubiquitous in eukaryotes and is a key enzyme in the regulation of the intracellular levels of adenosine, an important physiological effector of many cells and tissues. In this paper we report the cloning of cDNAs encoding adenosine kinase from both rat and human tissues. Two distinct forms of adenosine kinase mRNA were identified in human tissues. Sequence variation between the two forms is restricted to the extreme 5′-end of the adenosine kinase mRNA, including a portion of the coding region, and is consistent with differential splicing of a single transcriptional product. We have expressed both forms inE. coliand produced soluble active enzyme which catalyzes the phosphorylation of adenosine with high specific activityin vitroand is susceptible to known adenosine kinase inhibitors.

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    Abbreviations: AK, adenosine kinaseUTR, untranslated region; 5′-RACE, 5′-Rapid Amplification of cDNA Ends; PCR, Polymerase Chain Reaction; RT-PCR, reverse transcription followed by PCR; BHK, Baby Hampster Kidney; ADO, adenosine;

    ☆☆

    F. C. Neidhardt, Ed.

    1

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