Expression and rescuing of a cloned human tumour necrosis factor gene using an EBV-based shuttle cosmid vector.
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.
D002460:Cell Line; D003001:Cloning, Molecular; D003360:Cosmids; D005796:Genes; D005822:Genetic Vectors; D006023:Glycoproteins / Q000235:genetics*; D006131:Growth Inhibitors / Q000235:genetics*; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D014158:Transcription, Genetic; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha
D006023:Glycoproteins; D006131:Growth Inhibitors; D014409:Tumor Necrosis Factor-alpha