Gene transfer in lymphoid cells: Expression of the Thy-1.2 antigen by Thy-1.1 BW5147 lymphoma cells transfected with unfractionated cellular DNA

J. W. Berman, R. S. Basch, A. Pellicer

Research output: Contribution to journalArticle

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Abstract

We have transferred the gene coding for the Thy-1.2 alloantigen into a Thy-1.1-bearing T-cell lymphoma. BALB/c thymocyte DNA, precipitated with calcium phosphate, was used to effect the transfer. We report the stable transformation of lymphoid cells by total cellular DNA. To our knowledge, this has not been previously reported. Transient expression of the transfected gene could be detected by flow cytofluorometry, and 5% (range, 1.5%-11%) of the recipient cells had Thy-1.2 antigen detectable on their surface 48 hr after transfer. 'Stable' transformants were isolated by repeatedly selecting for cells expressing the Thy-1.2 antigen, by use of fluorescence-activated cell sorting or 'panning'. No metabolic selection was required. The transferred gene, detected by Southern blotting, encoded a product that is indistinguishable from the normal antigen by immunoprecipitation and NaDodSO4/PAGE.

Original languageEnglish (US)
Pages (from-to)7176-7179
Number of pages4
JournalProceedings of the National Academy of Sciences of the United States of America
Volume81
Issue number22 I
DOIs
StatePublished - Jan 1 1984
Externally publishedYes

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