Molecular cloning and primary structure of the avian Thy-1 glycoprotein.
We have previously characterised, both biochemically and immunohistochemically, a 23 kDa putative avian Thy-1 protein homologue. In this report we have examined the carbohydrates present on the protein and determined the partial protein sequence of enzymatically and CNBr-produced peptides. The protein sequences enabled us to clone an essentially full-length (1854 bp) cDNA using PCR and colony screening of an embryonic day (ED) 18 forebrain pUEX-1 cDNA library. Analysis of deduced amino acid sequence shows the 23 kDa protein to be 110 amino acids in length compared to mouse (112) and human and rat (111) while still retaining the conserved cysteine residues. The N-glycosylation site at position 61 is the same as that in the human protein, but is different from that in the rodent (position 75). Northern blot analysis of Thy-1 mRNA expression in the forebrain, cerebellum and tectum show that all three tissues have low levels at ED4 (forebrain and tectum) and ED8 (cerebellum), rising most rapidly between ED16 and the first few days post-hatch. Analysis of various tissues at hatch and adult show expression to be predominantly neuronal with very low levels in some other organs, mainly at hatch, indicating the possibility of subsets of cells, which we have also seen in histological sections, in these tissues expressing Thy-1 mRNA. Bone marrow and blood cells were also negative for Thy-1 protein.
Children's Medical Research Foundation, Camperdown, N.S.W. Australia.
B J
BJ
Dowsing
A A
AA
Gooley
P
P
Gunning
A
A
Cunningham
P L
PL
Jeffrey
D000375:Aging; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000235:genetics* / Q000302:isolation & purification; D001483:Base Sequence; D002645:Chickens; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D005314:Embryonic and Fetal Development; D015723:Gene Library; D006026:Glycoside Hydrolases; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D009928:Organ Specificity; D016133:Polymerase Chain Reaction; D016548:Prosencephalon / Q000196:embryology / Q000254:growth & development / Q000502:physiology*; D017385:Sequence Homology; D018800:Thy-1 Antigens
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D008562:Membrane Glycoproteins; D009838:Oligodeoxyribonucleotides; D018800:Thy-1 Antigens; D004247:DNA; D006026:Glycoside Hydrolases