Protoplast fusion method for high-frequency DNA transfection in human cells
US4608339A · kind A · utility
Inventors
Key dates
| Filing date | Oct 25, 1983 |
| Grant date | Aug 26, 1986 |
| Priority date | — |
| Expiry date | Oct 25, 2003 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N2730/10122
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A modified protoplast fusion method and cell line is disclosed that stably transfects human cells with pSV2-derived plasmids at frequencies greater than 10.sup.-3. This procedure makes it possible to test the biological effect of individual genes (i.e., oncogenes and other cellular genes, and viral genes). To demonstrate the utility of this invention, a pSV2gpt.sup.+ plasmid constructed to carry a subgenomic fragment of hepatitis B virus (HBV) that contained the core antigen gene (HBc gene) is transfected into human cells. Human cell lines are stably transfected with the HBC.sup.+ gene by selecting recipient cells for expression of guanine phosphoribosyl transferase expression; other selective markers, i.e., neomycin resistance, can also be used. Conditions for enhancing the expression of the transfected gene(s) have also been developed. For example, with this gpt.sup.+ /HBc.sup.+ cell line it is shown that growth in serum-free medium or treatment with 5'-azacytidine stimulates the production of the HBV core antigen.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.