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Nucleic Acids Research, 1983, Vol. 11, No. 13 4307-4323
© 1983

MOLECULAR BIOLOGY

Molecular cloning of human interleukin 2 cDNA and its expression in E. coli

Rene Devos*, Geert Plaetinck, Hilde Cheroutre, Guus Simons, Wim Degrave, Jan Tavernier, Erik Remaut and Walter Fiers

Laboratory of Molecular Biology, State University of Ghent Ledeganckstraat 35, B-9000, Ghent, Belgium

Received May 6, 1983. Accepted June 14, 1983.

A recombinant plasmid containing human interleukin 2 (IL2) cDNA was identified in a cDNA library constructed from mRNA derived from PHA-TPA induced splenocytes. Using this cDNA as a hybridization probe, a DNA fragment containing the IL2 gene was isolated from a collection of hybrid phages derived from human genomic DNA. A unique reading frame was identified from the nucleotide sequence derived from these plasmids coding for a polypeptide of 153 amino acids and containing a putative signal sequence of 20 amino acids. A mature polypeptide starting with either Met-Ala-Pro or Met-Pro was expressed in E. coli under control of the E. coli trap promoter or using a combination of the phage {lambda}PL. promoter and a ribosome binding site derived from phage Mu. The bacterial IL2 polypeptide had a molecular weight of 15,000 daltons and accounted for more than 10% of the total E. coli proteins in fully induced cells; it was biologically active in the T-cell specific DNA synthesis assay, even after recovery from a SDS-containing polyacrylamide gel.

*Present address: Biogent, Plateaustraat 22, B-9000, Ghent, Belgium


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