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. 1987 Feb;6(2):355–361. doi: 10.1002/j.1460-2075.1987.tb04762.x

Expression and rescuing of a cloned human tumour necrosis factor gene using an EBV-based shuttle cosmid vector.

D Kioussis, F Wilson, C Daniels, C Leveton, J Taverne, J H Playfair
PMCID: PMC553403  PMID: 3034585

Abstract

A cosmid vector carrying the Epstein-Barr virus origin of replication, the EBNA-1 gene, the hygromycin phosphotransferase (hph) gene and pBR322 sequences has been constructed. This cosmid can replicate autonomously in the nucleus of human tissue culture cells, even when it carries a 35-kb long insert. The cosmid can be rescued from the transfected cells by cloning it directly into ampicillin-sensitive Escherichia coli. A gene for human tumour necrosis factor (TNF) cloned into this cosmid vector was introduced in tissue culture cells, where it was transcribed into mature mRNA.

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