Molecular Cloning and Expression of Cynomolgus Monkey Interleukin -1β cDNA.

Autor: Totsuka, Kazuya, Takakura, Haruka, Hashimoto, Osamu, Tatsumi, Masashi
Předmět:
Zdroj: International Archives of Allergy & Immunology; May1998, Vol. 116 Issue 1, p40-48, 9p
Abstrakt: The cynomolgus monkey cDNA encoding interleukin-1β (IL-1β) was molecularly cloned by the reverse transcription polymerase chain reaction from a cDNA library of adherent splenocytes stimulated with lipopolysaccharides. The sequence analysis showed that the monkey IL-1β cDNA encodes a protein of 268 amino acids and displays a high degree (90%) of homology with the human counterpart. Substitution of amino acids resides mainly in the leader sequences of IL-1β when compared with those of human IL-1β. This cloned monkey IL-1β cDNA was used to express in Escherichia coli as a fusion protein with thioredoxin and in insect cells infected with a recombinant baculovirus after molecular modification where monkey IL-1β signal sequences were placed prior to the mature sequences of IL-1β for efficient secretion in insect cells. Recombinant monkey IL-1β expressed in both systems was shown to react with rabbit antihuman IL-1β antiserum by Western blot analysis and to have the biological activity of IL-1β in a bioassay. [ABSTRACT FROM AUTHOR]
Databáze: Complementary Index