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. 2001 Apr 10;98(9):5031–5036. doi: 10.1073/pnas.091097298

Figure 8.

Figure 8

(Upper) Copy number determination of the integrated lacZ gene. Lanes: 1, parent cell line 293 M+; 2, mutant 5-MCDG; 3, cell line with empty vector; 4, cell line with 5-MCDG. Total genomic DNA was digested with EcoRI. EcoRI cuts only once in the integrated construct with a second cut generating the 6-kb fragment in surrounding genomic DNA. In Southern blots, the coding region of lacZ was used as a labeled probe. (Lower) Under the antibiotic selection of transformants, there is no amplification of the reporter gene. Slot blots of total genomic DNA (cleaved with EcoRI) where the coding region of lacZ was used as labeled probe. Rows: A, cell line with 5-MCDG; B, cell line with the empty vector; C, cell line with the mutant 5-MCDG; D, parent cell line 293 M+. 293 M+ is the parent cell line and is transfected with the mutant of 5-MCDG, the empty vector, or 5-MCDG.