The Journal of Cell Biology, Vol 86, 341-346, Copyright © 1980 by The Rockefeller University Press
DNA-mediated cotransfer of unlinked mammalian cell markers into mouse L cells
H Warrick, N Hsiung, TB Shows and R Kucherlapati
Purified DNA from three different types of mammalian cells was precipitated
with calcium phosphate and added to mouse L cells deficient in thymidine
kinase (TK). Donor DNA was prepared from three cell lines: (a) mouse cells
transfected with UV-inactivated herpes simplex virus (HSV) type 1, or a
purified fragment of HSV carrying the TK gene (b) human HeLa cells, and (c(
CHO, a cell line derived from Chinese hamster ovaries. Several
hypoxanthine-aminopterin-thymidine resistant colonies were isolated from
each experiment. The origin of the TK that is expressed in these cells was
studied by polyacrylamide gel electrohporesis, isoelectric focusing, or
heat stability. The TK in all instances was of the donor origin. To
determine the extent of gene transfer we have assayed the CHO and HeLa DNA
transfectants for galactokinase (GALK), a marker closely linked to TK, and
25 other isozymes representing a large number of different chromosomes. No
cotransfer of GALK was observed, indicating that the size of the
transferred DNA segment is limited. We observed that, in one instance,
esterase-D, an unlinked marker of Chinese hamster origin, was transferred
along with TK. These experiments indicate that nonselected markers can be
transferred by this method, although at a low efficiency.
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