Molecular Determinants of Antiestrogen and Drug Sensitivity in Breast Carcinoma Cells
Abstract
Over the past year, we have continued the optimization of the cell/vector system and the procedures for the selection of genetic suppressor elements (GSEs) conferring drug resistance in MCF7 breast carcinoma cells. We have developed a rapid and efficient procedure for the recovery of integrated retroviruses from the DNA of selected cells. We have also identified retroviral vector LmGCX, which carries green fluorescent protein (GFP) as a selectable marker, as an efficient vehicle for gene expression and GFP selection in MCF7 cells. The LmGCX vector was used to generate a library of normalized random cDNA fragments from MCF7 cells. This library includes 29 x 10(exp 8) clones, >90% of which are recombinant. The cDNA fragments in this library carry different adaptors at their 5' and 3' termini (as oriented relative to the original mRNA), providing translation initiation and termination codons for peptide expression. This library has been used to initiate the selection of GSEs conferring resistance to Adriamycin and methotrexate in MCF7 cells.
Document Details
- Document Type
- Technical Report
- Publication Date
- Aug 01, 1997
- Accession Number
- ADB232313
Entities
People
- Igor B Roninson
Organizations
- University of Illinois at Chicago