Transient transfection of mammalian cells with DNA of the plant-pathogenic Ti-plasmid and expression of marker and resident sequences.
The large Ti-plasmid from Agrobacterium tumefaciens strain C58 has been used for transfection experiments with mammalian cells. In DNA from Tupaia baby fibroblasts Ti-plasmid sequences could be identified by filter hybridization as long as four weeks after transfection including two cell passages. The hybridization signals decreased rapidly after addition of the Ti-plasmid DNA-coprecipitate to the cells. The signals were often not detected any more after the first day, but were visible one week after transfection. Nuclei prepared from Ti-plasmid-transfected cells hybridized to pTi-specific RNA. With the chloramphenicol acetyl transferase-gene as marker no discrimination in DNA uptake was found between the Ti-plasmid and much smaller plasmids. According to the number of nuclei with homology to pTi-sequences it is assumed that about 0.2% of the cells carry Ti-plasmid DNA in the nucleus. Analysis of RNA isolated from cells transfected with cloned segments of the Ti-plasmid revealed that the T-DNA region of the Ti-plasmid was predominantly transcribed.
Geider
K
K
Max-Planck-Institut für medizinische Forschung, Abteilung Molekulare Biologie, Heidelberg, FRG.
Darai
G
G
Rhim
S L
SL
Simon
H G
HG
Moos
M
M
Harth
G
G
D000818:Animals; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics*; D005796:Genes; D005798:Genes, Bacterial; D009693:Nucleic Acid Hybridization; D010944:Plants / Q000382:microbiology*; D010957:Plasmids; D012329:RNA, Bacterial / Q000235:genetics; D012231:Rhizobium / Q000235:genetics*; D014162:Transfection
D004269:DNA, Bacterial; D012329:RNA, Bacterial; D015500:Chloramphenicol O-Acetyltransferase