Cross-species PCR cloning of gerbil (Meriones unguiculatus) interleukin-2 cDNA and its expression in COS-7 cells.
A cDNA clone of the gerbil interleukin-2 (IL-2) gene was isolated by cross-species PCR cloning, and demonstrated to produce a functional gerbil IL-2 protein when inserted in the eucaryotic expression vector pSV-SPORT1 and transfected into COS-7 monkey cells. The open reading frame codes for a polypeptide of 155 amino acid residues with a molecular weight (MW) of 17,601 which includes a putative signal peptide. The mature gerbil IL-2 is deduced to contain 135 amino acid residues and has a calculated MW of 15,496. Culture supernatant of COS-7 cells transfected with pSV-SPORT1-GIL-2, but not pSV-SPORT1 stimulates the proliferation of the IL-2 dependent murine CTLL-2 cells. Molecular characteristics of gerbil IL-2 have been compared with IL-2 of mouse, rat, human, bovine, ovine and porcine origin. The mature form of gerbil IL-2 is similar in molecular weight to all species except the mouse. A N-glycosylation site present in bovine, ovine and porcine IL-2 respectively, is absent in gerbil. Three Cys residues are conserved in all compared mature IL-2 molecules. In these comparisons, gerbil IL-2 has highest identity with rat IL-2 for both nucleotide and amino acid sequence.
Mai
Z
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Department of Veterinary Microbiology and Parasitology, School of Veterinary Medicine, Louisiana State University, Baton Rouge 70803.
Kousoulas
K G
KG
Horohov
D W
DW
Klei
T R
TR
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002460:Cell Line; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004247:DNA; D005849:Gerbillinae / Q000235:genetics*; D006801:Humans; D007376:Interleukin-2 / Q000235:genetics* / Q000276:immunology; D008251:Macaca; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D017386:Sequence Homology, Amino Acid; D012756:Sheep; D013552:Swine; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014162:Transfection
D007376:Interleukin-2; D011994:Recombinant Proteins; D004247:DNA