| Autor: |
de Saint Vincent BR, Delbrück S, Eckhart W, Meinkoth J, Vitto L, Wahl G |
| Jazyk: |
angličtina |
| Zdroj: |
Cell [Cell] 1981 Dec; Vol. 27 (2 Pt 1), pp. 267-77. |
| DOI: |
10.1016/0092-8674(81)90410-4 |
| Abstrakt: |
Rodent cells resistant to PALA, a specific inhibitor of the aspartate transcarbamylase activity of the multifunctional CAD protein, overproduce CAD as a result of amplification of the CAD gene. We cloned a functional CAD gene from Syrian hamster cells using a cosmid vector. Two independently isolated cosmids containing CAD genes have inserts 40 and 45 kb long. We introduced the cloned genes into CAD-deficient Chinese hamster ovary (CHO) cell mutants by fusing them with protoplasts of Escherichia coli containing the cosmids. We also introduced the cloned genes into wild-type CHO cells by selecting cells that became resistant to high concentrations of PALA following protoplast fusion. The transformants of the mutant and wild-type CHO cells contain multiple active copies of the donated Syrian hamster CAD genes. The cloned genes in three independent transformants are integrated into host-cell chromosomes at single locations identified by in situ hybridization. In two of these transformants, the genes are located in one X chromosome or in a chromosome resembling the X. In the third case, the genes are located in a small metacentric or rearranged chromosome. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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