EBV-based plasmid DNA rearrangements after transfection of eukaryotic cells.
The cDNA encoding influenza virus (A/Udorn/307/72 strain) M2 protein was subcloned into the EBV-based vector pREP9. Three continuous kidney cellular lines of different origin were transfected with recombinant plasmid pREP9-M2. One and 5 months after transfection plasmid DNA rearrangements were detected by means of restriction analysis of recovered plasmids and their hybridization with an influenza-virus-specific radioactive probe. Deletions were the most frequent type of pREP9-M2 mutations. PCR with primers corresponding to cellular genome and plasmid DNA followed by Southern blot analysis with the [(32)P]-labeled M2-fragment allowed host DNA rearrangements to be revealed in transfected cells.
Morozova
O V
OV
Novosibirsk Institute of Bioorganic Chemistry, Lavrentyev's Prospect 8, Novosibirsk, 630090, Russia.
Maksimova
T G
TG
Kostenko
E V
EV
D000818:Animals; D015139:Blotting, Southern; D002460:Cell Line; D015247:Deoxyribonuclease HindIII / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D004285:Dogs; D015321:Gene Rearrangement; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D007668:Kidney / Q000166:cytology; D010957:Plasmids / Q000235:genetics*; D015183:Restriction Mapping; D013552:Swine; D014162:Transfection; D014763:Viral Matrix Proteins / Q000235:genetics; D000086082:Viroporin Proteins
D015247:Deoxyribonuclease HindIII; D015252:Deoxyribonucleases, Type II Site-Specific; D014763:Viral Matrix Proteins; C031540:M-protein, influenza virus; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins; C029312:GGTACC-specific type II deoxyribonucleases