Premium
Transfer of the Epstein‐Barr virus genes coding for small RNAs to human lymphoid cells with a vector carrying a dominant selectable marker.
Author(s) -
Rymo L.
Publication year - 1983
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1002/j.1460-2075.1983.tb01511.x
Subject(s) - biology , microbiology and biotechnology , ecori , gene , restriction enzyme , southern blot , dna , selectable marker , rna , epstein–barr virus , shuttle vector , vector (molecular biology) , genetics , virus , transformation (genetics) , recombinant dna
Epstein‐Barr virus (EBV)‐negative, Burkitt‐like lymphoma‐derived cells were transformed with a transducing vector (pSV2‐gpt) containing the Escherichia coli gene coding for xanthine‐guanine phosphoribosyltransferase (XGPRT) and with a derivative of PSV2‐gpt that carries the genes for the EBV‐associated small RNAs on the EcoRI J fragment of B95‐8 EBV DNA inserted at the unique EcoRI site (pJ‐gpt). Cells transformed with PSV2‐gpt and pJ‐gpt express the E. coli gpt gene to approximately the same extent, judged by determinations of the XGPRT activity of cell extracts. Blot hybridisation experiments with restriction endonuclease‐cleaved DNA from the transformants have revealed the presence of vector DNA sequences in the cells, at least some of which are most probably integrated into high mol. wt. chromosomal DNA. Northern blot hybridisation analysis of cytoplasmic RNA from pJ‐gpt‐transformed cells revealed the presence of an EcoRI J DNA complementary RNA species of the same size as the EBV DNA‐encoded small RNAs found in EBV‐transformed cells.