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. 1984 Nov;4(11):2549–2552. doi: 10.1128/mcb.4.11.2549

Efficient transfer of cloned DNA into human diploid cells: protoplast fusion in suspension.

P Litzkas, K K Jha, H L Ozer
PMCID: PMC369089  PMID: 6096698

Abstract

A method for fusion of protoplasts bearing amplified plasmids and human diploid fibroblasts or other cell types in suspension is described. Transient expression of plasmid-encoded proteins occurs in up to 50% of the human cells, as demonstrated for simian virus 40 T antigen by immunofluorescence and the Escherichia coli xanthine-guanine phosphoribosyl transferase by autoradiography. In contrast, frequencies of stable transformants were similar to those obtained by the CaPO4 coprecipitation technique. However, experiments with both methods involving the recombinant pRSVneo (in which the Rous sarcoma virus long terminal repeat regulates expression of the antibiotic-inactivating aminoglycoside phosphotransferase) revealed a much higher frequency of colonies in G418 selective medium with constructions in which the early region of simian virus 40 DNA was present as well. We propose a role for the simian virus 40 T antigen in enhancing stable transformation in this system.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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