Isolation and characterization of cDNA clones for mouse Ly-9.
We describe the production and characterization of a mouse mAb, S-450-33.2, recognizing the Ly-9.2 specificity. This mAb was used to purify Ly-9 molecules from lymphoid cell lines, and the amino-terminal amino acids were determined. The mAb was also used in a eukaryotic expression system, to isolate cDNA clones encoding Ly-9. Analysis of RNA showed that Ly-9 expression is lymphocyte specific, as determined by the presence of a single hybridizing 2.4-kb species found only in lymphoid cells. Genomic DNA analysis indicated that Ly-9 is encoded by a single-copy gene of 10 to 15 kb. The predicted polypeptide belongs to the Ig superfamily of cell surface molecules with four extracellular Ig-like domains, i.e., a non-disulfide-bonded V domain, a truncated C2 domain with two disulfide bonds, a second non-disulfide-bonded V domain, and a truncated C2 domain with two disulfide bonds (V-C2-V-C2). The sequence data also support the view that Ly-9 belongs to the subgroup of the Ig superfamily that includes Bcm-1, CD2, and LFA-3.
Sandrin
M S
MS
Molecular Immunogenetics Laboratory, Austin Research Institute, Austin Hospital Heidelberg, Victoria, Australia.
Gumley
T P
TP
Henning
M M
MM
Vaughan
H A
HA
Gonez
L J
LJ
Trapani
J A
JA
McKenzie
I F
IF
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000950:Antigens, Ly / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification*; D005803:Genes, Immunoglobulin; D051379:Mice; D008969:Molecular Sequence Data; D014162:Transfection
D000911:Antibodies, Monoclonal; D000950:Antigens, Ly; D004247:DNA