Molecular cloning and expression of cynomolgus monkey interleukin-1beta cDNA.
The cynomolgus monkey cDNA encoding interleukin-1beta (IL-1beta) 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-1beta 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-1beta when compared with those of human IL-1beta. This cloned monkey IL-1beta 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-1beta signal sequences were placed prior to the mature sequences of IL-1beta for efficient secretion in insect cells. Recombinant monkey IL-1beta expressed in both systems was shown to react with rabbit antihuman IL-1beta antiserum by Western blot analysis and to have the biological activity of IL-1beta in a bioassay.
Totsuka
K
K
Department of Veterinary Science, National Institute of Infectious Diseases, Tokyo, Japan.
Takakura
H
H
Hashimoto
O
O
Tatsumi
M
M
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D007313:Insecta / Q000166:cytology / Q000235:genetics; D007375:Interleukin-1 / Q000235:genetics*; D008252:Macaca fascicularis / Q000235:genetics*; D008969:Molecular Sequence Data; D011506:Proteins / Q000032:analysis / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D013879:Thioredoxins / Q000235:genetics; D014162:Transfection / Q000235:genetics
D018076:DNA, Complementary; D007375:Interleukin-1; D011506:Proteins; D011993:Recombinant Fusion Proteins; D013879:Thioredoxins