vector permitting positive selection for integration
Comments
Restriction digests of the clone give the following sizes (kb): BamHI--8.6, 1.36; EcoRI--9.4; HindIII--9.4.
Transfected methotrexate-resistant NIH 3T3 cells carried double minute chromosomes that appeared to contain DNA acquired during transfection.
Can be used in cotransfection experiments to generate a selectable methotrexate-resistant phenotype.
A 3.3 kb EcoRI/BamHI fragment of HaSV clone H-1 (including the LTR) was cloned into EcoRI/BamHI-digested pBR322. A PstI fragment with mouse Dhfr (with BamHI linkers) was then cloned into the BamHI site of the construct.
Media
ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References
Murray MJ, et al. Construction and use of a dominant, selectable marker: a Harvey sarcoma virus-dihydrofolate reductase chimera. Mol. Cell. Biol. 3: 32-43, 1983. PubMed: 6298606