Homologous recombination of SV40 DNA in COS7 cells occurs with high frequency in a gene dose independent fashion.
Homologous recombination between microinjected SV40 DNA fragments and endogenous SV40 DNA in COS7 cells was analysed by immunofluorescence staining and DNA blotting. Time course experiments revealed that recombination between the transferred (trans) DNA and the chromosomal DNA occurred about 8 hours after microinjection with high efficiency in a gene dose independent fashion. Deletions of up to 1018 basepairs (bp) within the early or the late SV40 region were efficiently repaired after the transfer of linear but not of circular DNA molecules. A 22 bp homology between the trans DNA and the endogenous DNA was sufficient to initiate recombination but 14 nonhomologous bp at one open end of the SV40 DNA fragments hindered gap repair.
Rommerskirch
W
W
Institut für Molekularbiologie und Biochemie, Freie Universität Berlin, FRG.
Graeber
I
I
Grässmann
M
M
Grässmann
A
A
D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D004270:DNA, Circular; D004274:DNA, Recombinant; D004279:DNA, Viral / Q000378:metabolism*; D005347:Fibroblasts; D008845:Microinjections; D011995:Recombination, Genetic; D012689:Sequence Homology, Nucleic Acid; D013539:Simian virus 40 / Q000235:genetics*
D004270:DNA, Circular; D004274:DNA, Recombinant; D004279:DNA, Viral