Molecular cloning of gerbil interleukin 12 and its expression as a bioactive single-chain protein.
Complementary DNAs coding for gerbil interleukin 12 (IL-12) p40/p35 subunits were cloned by a combination of cross species reverse transcriptase-polymerase chain reaction (RT-PCR) and 3' rapid amplification of cDNA ends (RACE) techniques. IL-12 p40/p35 had 79% nucleotide identity and 81% amino acid homology to mouse IL-12 p40/p35. The p40/p35 subunits were expressed as a single polypeptide separated by a short hinge sequence that allowed for proper folding and assembly. COS-7 cells transfected with DNA encoding the single-chain gerbil IL-12 (pSCjIL12) secreted high levels of the protein which stimulated proliferation of ConA-activated gerbil spleen lymphoblasts in a dose-dependent manner.
Gaucher
D
D
Institute of Parasitology, McGill University, Macdonald Campus, 21,111 Lakeshore Road, Ste. Anne de Bellevue, Quebec, Canada H9X 3V9.
Chadee
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002460:Cell Line; D003001:Cloning, Molecular; D005260:Female; D005849:Gerbillinae / Q000235:genetics / Q000276:immunology*; D018664:Interleukin-12 / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D008213:Lymphocyte Activation; D051379:Mice; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D014162:Transfection
D018664:Interleukin-12