THE HUMAN MCP-2 GENE (SCYA8) - CLONING, SEQUENCE-ANALYSIS, TISSUE EXPRESSION, AND ASSIGNMENT TO THE CC CHEMOKINE GENE CONTIG ON CHROMOSOME-17Q11.2

Monocyte chemotactic proteins (MCPs) form a sub-family of chemokines t hat recruit leukocytes to sites of inflammation and that may contribut e to tumor-associated leukocyte infiltration and to the antiviral stat e against HIV infection. With the use of degenerate primers that were based on CC chemokine consensus sequences, the known MIP-1 alpha/LD78 alpha, MCP-1, and MCP-3 genes and the previously unidentified eotaxin and MCP-2 genes were isolated from a YAC contig from human chromosome 17q11.2. The amplified genomic MCP-2 fragment was used to isolate an M CP-S cosmid from which the gene sequence was determined. The MCP-2 gen e shares with the MCP-1 and MCP-3 genes a conserved intron-exon struct ure and a coding nucleotide sequence homology of 77%. By Northern blot analysis the 1.0-kb MCP-2 mRNA was predominantly detectable in the sm all intestine, peripheral blood, heart, placenta, lung, skeletal muscl e, ovary, colon, spinal cord, pancreas, and thymus. Transcripts of 1.5 and 2.4 kb were found in the testis, the small intestine, and the col on. The isolation of the MCP-2 gene from the chemokine contig localize d it on YAC clones of chromosome 17q11.2, which also contain the eotax in, MCP-1, MCP-3, and NCC-1/MCP-4 genes. The combination of using dege nerate primer PCR and YACs illustrates that novel genes can efficientl y be isolated from gene cluster contigs with less redundancy and effor t than the isolation of novel ESTs. (C) 1997 Academic Press.