Cloning and characterization of ovine beta2-microglobulin cDNAs.
Beta-2-microglobulin (beta(2)m) is the light chain of the major histocompatibility complex (MHC) class I cell surface heterodimer. beta(2)m is well conserved across most species with few polymorphisms seen within species. The aims of this study were to clone and express ovine beta(2)m and investigate if allelic variation of ovine beta(2)m exists. Ovine beta(2)m clones were isolated from five sheep of three breeds by reverse-transcription polymerase chain reaction (RT-PCR). Sequence analysis showed that four ovine beta(2)m sequences were obtained. Within breeds and individual animals there was evidence of allelic variation of ovine beta(2)m. An expression system was established to express one of the alleles with an ovine MHC class I cDNA clone in human embryo kidney cells (HEK293) and quail cells (QT35). Transfection experiments showed that ovine beta(2)m was expressed and directed the expression of ovine MHC class I heavy chain to the cell surface of the transfected cells. Both bovine and human beta(2)m supported ovine MHC class I heavy chain cell surface expression.
Wu
Changxin
C
Department of Veterinary Medicine, University of Cambridge, Madingley Road, Cambridge CB3 0ES, UK. cxw20@cam.ac.uk
McConnell
Ian
I
Blacklaws
Barbara
B
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005434:Flow Cytometry / Q000662:veterinary; D008969:Molecular Sequence Data; D012313:RNA / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D016415:Sequence Alignment; D012756:Sheep / Q000235:genetics* / Q000276:immunology; D014162:Transfection / Q000662:veterinary; D001613:beta 2-Microglobulin / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology
D018076:DNA, Complementary; D001613:beta 2-Microglobulin; D012313:RNA