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Effective synthesis and cloning of the human interleukin-2 gene and its analogue; expression of interleukin-2 in E. coli
N. K. Danilyuk, S. V. Seryogin, A. N. Sinyakov, O. I. Serpinski, I. N. Babkina, M. A. Urmanova, V. A. Ryabinin, S. G. Pozdnyakov
All-Union Research Institute of Molecular Biology, Koltsovo, Novosibirsk Region
Abstract: Artificial DNA fragments encoding human interleukin-2 (133 a.a.) and its analogue (deletion of 14 C-terminal a.a.) were prepared by means of the DNA polymerase I mediated extention of synthetic polynucleotides having short overlapping sequences at their 3'-ends. The fragments were cloned in specially designed pFH-type plasmids and then excised by the FokI and other restriction endonucleases to yield the sub-fragments with the structurally predetermined 5'-unique cohensive ends. The complete synthetic gene was constructed by one or two-step ligation. The expressed IL-2 was tested by analysing the T-cell proliferation activity of E.coli crude lisates containing the pEXIL2 expression plasmid.
Russian Journal of Bioorganic Chemistry 1991, 17 (6):779-788