Cloning and expression of a lymphocyte activation gene (LAG-1).
Using subtractive hybridization of a cDNA library we have identified a human gene, termed LAG-1 (for "Lymphocyte Activation Gene-1"). This cDNA codes for a 69 amino-acid polypeptide which belongs to a new class of recently described proteins secreted by activated lymphocytes and/or monocytes. The LAG-1 gene was cloned, sequenced and its chromosomal location assigned to chromosome 17 (q21 band). The promoter region of the LAG-1 gene was shown to include a GM-CSF-like decanucleotide sequence. Using a baculovirus vector expression system, we found that a 10 kDa recombinant LAG-1 protein is secreted by AcNPV infected SF9 cells, as determined in Western blot experiments by the reactivity of specific anti-peptidic heteroantibodies. Finally the natural LAG-1 protein was precipitated from the supernatant of internally labeled activated Nk cells and shown to migrate as a single entity of 14 kDa in SDS-PAGE analysis.
Baixeras
E
E
Laboratoire d'Immunologie Cellulaire, Institut Gustave-Roussy, France.
Roman-Roman
S
S
Jitsukawa
S
S
Genevee
C
C
Mechiche
S
S
Viegas-Pequignot
E
E
Hercend
T
T
Triebel
F
F
D000595:Amino Acid Sequence; D000906:Antibodies; D001402:B-Lymphocytes / Q000378:metabolism; D016367:Baculoviridae; D001483:Base Sequence; D015139:Blotting, Southern; D054407:Chemokine CCL4; D019742:Chemokines, CC; D002874:Chromosome Mapping; D002886:Chromosomes, Human, Pair 17; D003001:Cloning, Molecular; D015723:Gene Library; D015698:Genomic Library; D006801:Humans; D007694:Killer Cells, Natural / Q000378:metabolism; D008213:Lymphocyte Activation / Q000235:genetics*; D019402:Macrophage Inflammatory Proteins; D008969:Molecular Sequence Data; D009928:Organ Specificity; D011233:Precipitin Tests; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics* / Q000302:isolation & purification; D012333:RNA, Messenger / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic; D014162:Transfection
D000906:Antibodies; C066507:CCL4 protein, human; D054407:Chemokine CCL4; D019742:Chemokines, CC; D019402:Macrophage Inflammatory Proteins; D011506:Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins