(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.