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Status and transcriptional activity of a bovine‐papillomavirus‐I‐based expression vector in a recombinant production cell line
Author(s) -
MR Fibi,
Kunz J.,
Weimer T.,
Schulz R.,
Teifke P.,
KH Grzeschik,
Johannsen R.,
Zettlmeissl G.
Publication year - 1996
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1111/j.1470-8744.1996.tb00367.x
Subject(s) - biology , transcription (linguistics) , recombinant dna , bovine papillomavirus , expression vector , extrachromosomal dna , cell culture , vector (molecular biology) , promoter , microbiology and biotechnology , genome , transcription factor , genetics , gene , gene expression , philosophy , linguistics
We have analysed the status and transcriptional activity of the bovine papillomavirus‐I (complete BPV‐I genome)‐based expression vector pCES in CI27i‐cell‐line‐derived 3TI cells used for the industrial production of recombinant human erythropoietin (rhuEpo). Complete tandem head‐to‐tail integration of about 600 vector copies at a single site of the cellular genome was observed. Deletions, insertions or rearrangements of pCES‐specific sequences or extrachromosomal copies of vector sequences were not detected. Transcriptional analyses demonstrated that rhuEpo was abundantly expressed. BPV‐I early transcription was detected, as expected from a BPV‐I‐transformed cell line; however, compared with the mouse metallothionein‐I‐promoter‐driven rhuEpo‐specific transcription, it was very weak. Late BPV‐I transcription was also not detected in 3TI cells under conditions of large‐scale rhuEpo production. Therefore this expression system proved to be safe and suitable for the production of rhuEpo.