Isolation and characterization of S49 mouse lymphoma cell mutants deficient in adenosine deaminase

Teh sheng Chan, Cecilia Huang, Toshiaki Sato

Research output: Contribution to journalArticle

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Abstract

Adenosine deaminase-deficient mutants of a mouse lymphoma cell line S49 have been isolated by a two-step selection process. In the first step, we derived mutant lines containing haploid levels of adenosine deaminase activity from wild-type cells. The selective medium contained tritiated deoxyadenosine, deoxycytidine, and deoxycoformycin. Wild-type cells were killed, presumably because of suicidal incorporation of tritiated deoxy adenosine via the adenosine deaminase pathway. The second step was to derive, from the partially deficient mutants, sublines that were virtually lacking adenosine deaminase, using tritiated deoxyadenosine and deoxycytidine. Four mutant clones were found to contain less than 5% of the enzyme activity of wild-type cells and virtually no immunoreactive adenosine deaminase protein. Northern blot analysis showed that the levels of adenosine deaminase mRNA were drastically reduced. Back-selection for adenosine deaminase-positive revertants can be accomplished by using a medium containing deoxyadenosine (as a sole source of purine), aminopterin, and thymidine or, alternatively, by using deoxyadenosine alone in a serum-free medium.

Original languageEnglish (US)
Pages (from-to)411-420
Number of pages10
JournalSomatic Cell and Molecular Genetics
Volume15
Issue number5
DOIs
StatePublished - Sep 1989

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Adenosine Deaminase
Lymphoma
Deoxycytidine
Aminopterin
Pentostatin
Haploidy
Serum-Free Culture Media
Northern Blotting
Adenosine
Thymidine
Clone Cells
Cell Line
Messenger RNA
2'-deoxyadenosine
Enzymes

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

Isolation and characterization of S49 mouse lymphoma cell mutants deficient in adenosine deaminase. / Chan, Teh sheng; Huang, Cecilia; Sato, Toshiaki.

In: Somatic Cell and Molecular Genetics, Vol. 15, No. 5, 09.1989, p. 411-420.

Research output: Contribution to journalArticle

Chan, Teh sheng ; Huang, Cecilia ; Sato, Toshiaki. / Isolation and characterization of S49 mouse lymphoma cell mutants deficient in adenosine deaminase. In: Somatic Cell and Molecular Genetics. 1989 ; Vol. 15, No. 5. pp. 411-420.
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