Cloning of human adenosine deaminase cDNA and expression in mouse cells

Dinko Valeric; R. Scott McIvor; Steven R. Williams; Marja G.C. Duyvesteyn; Hans van Ormondt; Alex J. van der Eb; David W. Martin

doi:10.1016/0378-1119(84)90205-1

Cloning of human adenosine deaminase cDNA and expression in mouse cells

Dinko Valeric, R. Scott McIvor, Steven R. Williams, Marja G.C. Duyvesteyn, Hans van Ormondt, Alex J. van der Eb, David W. Martin

Research output: Contribution to journal › Article › peer-review

43 Scopus citations

Abstract

A previously isolated partial cDNA sequence encoding human adenosine deaminase (ADA) was used to probe a cDNA library prepared from human cultured cell mRNA. Clones containing a combined overlapping length of 1462 bp were isolated and sequenced. One of these was found to include the entire ADA coding region An open reading frame consisting of 363 codons was identified, predicting a polypeptide of M_r 40 762. A mammalian expression plasmid was constructed, positioning the ADA coding sequence to be under transcriptional control of the mouse metallothionein promoter. Transfection of cultured mouse L-cells with this plasmid resulted in the acute expression of human ADA enzymatic activity, as assayed by isoelectric focusing.

Original language	English (US)
Pages (from-to)	147-153
Number of pages	7
Journal	Gene
Volume	31
Issue number	1-3
DOIs	https://doi.org/10.1016/0378-1119(84)90205-1
State	Published - Nov 1984
Externally published	Yes

Bibliographical note

Funding Information:
We areg ratefutl o Albert Tam and Michael Shepard for supplyingt he mousem etallothioneipnr o-moter,t o Chris Simonsenfo r usefuld iscussiona nd to Loes van der Voorn for ablet echnicaal ssistance. This work was supportedb y a grantt o D.V. fromt he NetherlandsO rganizationf or the Advancemenot f Pure Research( ZWO) and by USPHS grant no. AM 20428f rom the National Instituteso f Health.

Copyright:
Copyright 2014 Elsevier B.V., All rights reserved.

Keywords

Recombinant DNA
eukaryotic expression vector
metallothionein promoter
sequencing
transfection

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title = "Cloning of human adenosine deaminase cDNA and expression in mouse cells",

abstract = "A previously isolated partial cDNA sequence encoding human adenosine deaminase (ADA) was used to probe a cDNA library prepared from human cultured cell mRNA. Clones containing a combined overlapping length of 1462 bp were isolated and sequenced. One of these was found to include the entire ADA coding region An open reading frame consisting of 363 codons was identified, predicting a polypeptide of Mr 40 762. A mammalian expression plasmid was constructed, positioning the ADA coding sequence to be under transcriptional control of the mouse metallothionein promoter. Transfection of cultured mouse L-cells with this plasmid resulted in the acute expression of human ADA enzymatic activity, as assayed by isoelectric focusing.",

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author = "Dinko Valeric and McIvor, {R. Scott} and Williams, {Steven R.} and Duyvesteyn, {Marja G.C.} and {van Ormondt}, Hans and {van der Eb}, {Alex J.} and Martin, {David W.}",

note = "Funding Information: We areg ratefutl o Albert Tam and Michael Shepard for supplyingt he mousem etallothioneipnr o-moter,t o Chris Simonsenfo r usefuld iscussiona nd to Loes van der Voorn for ablet echnicaal ssistance. This work was supportedb y a grantt o D.V. fromt he NetherlandsO rganizationf or the Advancemenot f Pure Research( ZWO) and by USPHS grant no. AM 20428f rom the National Instituteso f Health. Copyright: Copyright 2014 Elsevier B.V., All rights reserved.",

year = "1984",

month = nov,

doi = "10.1016/0378-1119(84)90205-1",

language = "English (US)",

volume = "31",

pages = "147--153",

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AU - Valeric, Dinko

AU - McIvor, R. Scott

AU - Williams, Steven R.

AU - Duyvesteyn, Marja G.C.

AU - van Ormondt, Hans

AU - van der Eb, Alex J.

AU - Martin, David W.

N1 - Funding Information: We areg ratefutl o Albert Tam and Michael Shepard for supplyingt he mousem etallothioneipnr o-moter,t o Chris Simonsenfo r usefuld iscussiona nd to Loes van der Voorn for ablet echnicaal ssistance. This work was supportedb y a grantt o D.V. fromt he NetherlandsO rganizationf or the Advancemenot f Pure Research( ZWO) and by USPHS grant no. AM 20428f rom the National Instituteso f Health. Copyright: Copyright 2014 Elsevier B.V., All rights reserved.

PY - 1984/11

Y1 - 1984/11

N2 - A previously isolated partial cDNA sequence encoding human adenosine deaminase (ADA) was used to probe a cDNA library prepared from human cultured cell mRNA. Clones containing a combined overlapping length of 1462 bp were isolated and sequenced. One of these was found to include the entire ADA coding region An open reading frame consisting of 363 codons was identified, predicting a polypeptide of Mr 40 762. A mammalian expression plasmid was constructed, positioning the ADA coding sequence to be under transcriptional control of the mouse metallothionein promoter. Transfection of cultured mouse L-cells with this plasmid resulted in the acute expression of human ADA enzymatic activity, as assayed by isoelectric focusing.

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KW - Recombinant DNA

KW - eukaryotic expression vector

KW - metallothionein promoter

KW - sequencing

KW - transfection

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Cloning of human adenosine deaminase cDNA and expression in mouse cells

Abstract

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Keywords

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