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

Please always quote using this URN: urn:nbn:de:bvb:20-opus-62662
  • A 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.A 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.show moreshow less

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Metadaten
Author: W. Lindenmaier, K. E. Dittmar, H. Hauser, A. Necker, Walter Sebald
URN:urn:nbn:de:bvb:20-opus-62662
Document Type:Journal article
Faculties:Fakultät für Biologie / Theodor-Boveri-Institut für Biowissenschaften
Language:English
Year of Completion:1985
Source:In: Gene (1985) 39(1) 33-39.
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie
GND Keyword:Biochemie
Tag:DNA mediated gene transfer; Recombinant DNA; bacteriophage lambda; expression plasmid; packaging; screening
Release Date:2012/04/26
Licence (German):License LogoDeutsches Urheberrecht