Isolation of a functional human interleukin 2 gene from a cosmid library by recombination in vivo.
A method has been developed that allows the isolation of genomic clones from a cosmid library by homologous 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 lambda phage particles. A recombination-proficient host strain carrying IL2 cDNA sequences in a non-homologous 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 gene was restored. After DNA mediated gene transfer into mouse Ltk- cells human IL2 was expressed constitutively.
D000818:Animals; D010582:Bacteriophage lambda / Q000235:genetics; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D003360:Cosmids; D004247:DNA / Q000235:genetics; D005796:Genes; D005822:Genetic Vectors; D006801:Humans; D007376:Interleukin-2 / Q000235:genetics*; D007739:L Cells / Q000378:metabolism; D051379:Mice; D011995:Recombination, Genetic
D007376:Interleukin-2; D004247:DNA