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Identification of the coding region for a second Epstein‐Barr virus nuclear antigen (EBNA 2) by transfection of cloned DNA fragments.
Author(s) -
MuellerLantzsch N.,
Lenoir G.M.,
Sauter M.,
Takaki K.,
Béchet J.M.,
KuklikRoos C.,
Wunderlich D.,
Bornkamm G.W.
Publication year - 1985
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.1460-2075.1985.tb03854.x
Subject(s) - biology , virology , transfection , microbiology and biotechnology , virus , dna , coding region , gammaherpesvirinae , plasmid , antigen , epstein–barr virus , gene , herpesviridae , genetics , viral disease
Cell lines were established by co‐transfection of cloned M‐ABA Epstein‐Barr virus (EBV) DNA fragments with plasmids conferring resistance to dominant selective markers. A baby hamster kidney cell line carrying the HindIII‐I1 fragment exhibits a nuclear antigen of 82 000 daltons, serologically defined as EBV‐determined nuclear antigen (EBNA) 1. Furthermore, a Rat‐1 cell line transfected with DNA of the clone pM 780‐28 containing three large internal repeats (BglII‐U) and the adjacent BglII‐C fragment expresses a nuclear antigen of 82 000 daltons which can be visualized only by a subset of anti EBNA‐positive human sera. Sera recognizing the 82 000‐dalton protein of the transfected cell line reacted with a protein of the same size in the non‐producer line Raji, designated as EBNA 2. Conversely, sera without reactivity to the 82 000‐dalton protein failed to react with EBNA 2 of Raji cells. P3HR‐1 and Daudi cells with large deletions in BglII‐U and ‐C are devoid of EBNA 2. The data presented provide evidence that a second EBNA protein is encoded by the region of the EBV genome which is deleted in the non‐transforming P3HR‐1 strain.