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phate co-precipitation to introduce the DNA into cultured cells--was not effi-
 

Summary: phate co-precipitation to introduce the
DNA into cultured cells--was not effi-
cient. With this method, incorporation
of functional copies of tk occurred in only one per million cells
exposed to the DNA­calcium phosphate co-precipitate. Using
a similar selection scheme, I sought to determine whether I
could introduce a functional tk into Tk­
cells using very fine
glass needles to inject DNA directly into nuclei34
. This proce-
dure proved extremely efficient. One cell in three that received
the DNA stably passed the functional tk to its daughter cells.
The high efficiency of DNA transfer by microinjection made it
practical for investigators to generate transgenic mice contain-
ing random insertions of exogenous DNA. This was accom-
plished by injection of the desired DNA into nuclei of one-cell
zygotes and allowing these embryos to come to term after sur-
gical transfer to foster mothers35­39
.
Efficient functional transfer of HSV-tk into cells required that

  

Source: Alvarado, Alejandro Sánchez - Department of Neurobiology and Anatomy, University of Utah

 

Collections: Biology and Medicine