The cloning and sequencing of cervine interleukin 10.
We report the cloning and sequencing of the cervine interleukin-10 gene. Specific cDNA was amplified by PCR using primers based on the bovine sequence. This was cloned into pGEM 5Zf and several clones were sequenced. The 762 nucleotide product coded for a 179 amino acid protein which was 86% homologous with its bovine and 77% homologous with its human counterparts. There is a strongly hydrophobic signal sequence consisting of the first 20 amino acids and a potential glycosylation site at amino acids 134-136. There are three regions, comprising 34% of the protein, which show complete homology between the cervine, bovine and human sequences. The transcription of the gene was shown by Northern Blotting where a single, 1.8kb, mRNA transcript was detected 4-8 hours after activation of peripheral blood mononuclear cells with mitogen.
Lockhart
E
E
Department of Microbiology, University of Otago, Dunedin, New Zealand.
Slobbe
L
L
Droogmans
L
L
Griffin
F
F
Buchan
G
G
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary; D003670:Deer / Q000235:genetics*; D016753:Interleukin-10 / Q000096:biosynthesis / Q000235:genetics*; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008969:Molecular Sequence Data; D011043:Pokeweed Mitogens / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D017386:Sequence Homology, Amino Acid; D013997:Time Factors
D017931:DNA Primers; D018076:DNA, Complementary; D011043:Pokeweed Mitogens; D012333:RNA, Messenger; D016753:Interleukin-10