Cloning of the gene for bovine monocyte chemoattractant protein-2.
Bovine monocyte chemoattractant protein-1 (bovine MCP-1) cDNA has recently been characterized and shown to be highly expressed in bovine seminal vesicles secretory epithelium as well as in phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear leukocytes (PMNLs). In an attempt to isolate the MCP-1 gene, we screened a bovine genomic cosmid library with a MCP-1-specific probe pH42. A positive clone, c11/1, was subjected to restriction analysis and fragments probed with pH42 by southern blotting. pH42-positive fragments were subcloned and sequenced. The sequence revealed three exon-like regions that coded for a protein displaying an identity of 51% with bovine MCP-1. Employing this sequence information from c11/1, the c11/1-specific cDNA was generated from poly(A)+RNA of bovine PMNLs by reverse transcription and a combination of polymerase chain reaction (PCR) methods. The assembled c11/1 cDNA comprised a 5' UTR coding region as well as 3' UTR for the gene product c11/1. Amino acid sequence comparison of the bovine c11/1 gene product with human monocyte chemotactic proteins yielded the highest sequence identity with human MCP-2, and it is assumed that the c11/1 gene product represents the bovine MCP-2. The exon/intron structure of the bovine MCP-2 gene was found to be similar to the human MCP-1 gene. The bovine MCP-2 gene consists of three exons separated by two introns. In the 5'-flanking region of the 3.3-kb gene, a TATA box as well as an AP-1 sequence motif were identified. The bovine MCP-2 is specified by a single-copy gene.(ABSTRACT TRUNCATED AT 250 WORDS)
Wempe
F
F
Laboratory of Genetic Engineering, Slovak Academy of Science, Bratislava, CSFR.
Hanes
J
J
Scheit
K H
KH
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D054412:Chemokine CCL8; D002630:Chemotactic Factors / Q000235:genetics*; D003001:Cloning, Molecular; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary; D015870:Gene Expression; D005796:Genes; D007438:Introns; D008969:Molecular Sequence Data; D018945:Monocyte Chemoattractant Proteins; D009000:Monocytes / Q000737:chemistry*; D012333:RNA, Messenger / Q000235:genetics; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
C515453:CCL8 protein, human; D054412:Chemokine CCL8; D002630:Chemotactic Factors; D017931:DNA Primers; D018076:DNA, Complementary; D018945:Monocyte Chemoattractant Proteins; D012333:RNA, Messenger