Isolation of a functional human interleukin 2 gene from a cosmid library by recombination in vivo

Abstract

SUMMARYA method has been developed that allows the isolation of genomic clones from a cosmid library by homologaus recombination in vivo. This method was used to isolate a human genomic interleukin 2 (IL2) gene. The genomic cosmid library was packaged in vivo into A. phage particles. A recombination-proficient host strain carrying IL2 cDNA sequences in a non-homologaus plasmid vector was infected by the packaged cosmid library. After in vivo packaging and reinfection, recombinants carrying the antibiotic resistance genes of both vectors were selected. From a recombinant cosmid clone the chromosomal IL2 genewas restored. After DNA mediated gene transfer into mouse Ltk-cells human IL2 was expressed constitutively. JNTRODUCTION

Cloning and characterization of major antigenic determinants of human cytomegalovirus Ad169 seen by the human immune system

“…3B, C, and data not shown). Clones from the subgenomic libraries were mapped by colony hybridization with subfragments of pCM1007 cloned in pAN26 [19], a Rl-derived vector without homologies to pEX1.35 clones of these ORF-libraries were analyzed in detail. All clones which were strongly reactive with human sera were clustered in the HindIII b, c, L region which had been shown to encode the 65 k "major late antigen" and a 71 k phosphoprotein [30,33].…”

Variation of amino acid concentrations in the medium of HU ?-IFN and HU IL-2 producing cell lines

“…They have been manipulated by genetic methods to produce human 13-interferon (Hauser et al, 1982) and human interleukin 2 (Lindenmaier et al, 1985) constitutively. Each cell line was cultivated in a 1 1 repeated batch fermentation process with a bubble free cell culture aeration system with porous moving membranes (Lehmann et al, 1985).…”