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The purpose of this investigation was to characterize the gene that encodes the receptor for mouse interferon-γ (IFN-γR), including determination of its size, intronic boundaries and its transcription start points (tsp). The mouseIFN -γR gene is 22-kb long, with six introns that range in size from approx. 1 to 7 kb. The first six exons encode the extracellular and transmembrane (TM) domains of the...
Phage display of proteins can be used to study ligand-receptor interaction and for the affinity-maturation of binding sites in polypeptide hormones and/or cytokines. We have expressed human interleukin-6 (hIL-6) on M13 phage in a monovalent fashion as a fusion protein with the phage coat protein, pIII. Phage-displayed hIL-6 is correctly folded, as judged by its ability to interact with conformation-specific...
The gene encoding the mouse cytotoxic T-lymphocyte-associated antigen 8 (CTLA8) has been cloned and its complete nucleotide (nt) sequence determined. Sequence and polymerase chain reaction (PCR) analysis indicated that the publishedCTLA8 sequence [Rouvier et al., J. Immunol. 150 (1993), 5445-5456] was of rat rather than mouse origin. The mouseCTLA8 gene contains two exons and one intron. The 5...
The nucleotide (nt) sequence encoding the bovine homologue of interleukin-5 (boIL-5) was determined. Total cDNA generated from stimulated bovine peripheral blood lymphocytes was used to amplify the boIL-5 gene using primers based on the 5 and 3 untranslated regions of the ovine IL-5 cDNA sequence. The boIL-5 coding sequence is 405 bp long, coding for a 15.2 kDa precursor protein of 134 amino...
Using lipopolysaccharide (LPS)-stimulated equine peripheral blood mononuclear cell (PBMC) cDNA as a template, we performed polymerase chain reaction (PCR) amplification with equine interleukin-1β (IL-1β) specific primers. Electrophoresis of the PCR product on agarose gel revealed an additional smaller fragment that hybridized with an equine IL-1β cDNA probe. Sequencing of this fragment demonstrated...
The MART-1/Melan-A human melanoma tumor antigen can be recognized by T lymphocytes and appears to be involved in tumor regression. To study the transcriptional regulation of this important gene, the 5 untranslated (UT) region of the MART-1/Melan-A gene was cloned and sequenced. Human melanoma cell lines were screened for MART-1/Melan-A mRNA expression. Primer extension and northern analysis were...
Interleukin-15 (IL-15) is a recently identified growth and differentiation factor with an important potential role in the initial immune responses to infection. To enable the study of the role of this cytokine in the protective immune-mechanisms generated against parasitic diseases of swine, cDNA was generated from a macrophage enriched adherent cell population from peripheral blood mononuclear cells...
Granulocyte colony-stimulating factor (G-CSF) is a cytokine that stimulates the proliferation and differentiation of hematopoietic progenitor cells committed to the neutrophil/granulocyte lineage. Recombinant G-CSF (rG-CSF) is routinely used in the prevention of chemotherapy-induced neutropenia and in the setting of bone marrow transplantation. Chronic idiopathic and congenital neutropenic disorders...
The sequence of the chicken interferon-γ (ifn-γ) gene was determined, one of the first non-mammalian cytokine gene structures to be elucidated. Initial genomic clones were amplified from chicken genomic DNA and were used to isolate a cosmid clone covering the entire gene for sequencing. The exon:intron structure of chicken ifn-γ is very similar to those of its mammalian homologues, with the exception...
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