Physiology and Endocrinology: Reproductive Technologies and Management

RPCI-42 Bovine Bacterial Artificial Chromosome (BAC) library was screened with oligonucleotide sequences corresponding to specific elements of the 5 UTR sequence of bovine PC and to a region of the coding sequence. Two BACs that hybridized to all probes were selected for further analysis. A partial restriction map of the BACs was made with oligonucleotides corresponding to the coding region and the 89 and 110 bp elements of bovine PC 5 UTR. The BAC fragments that hybridized to the oligonucleotide probes were isolated and sequenced. The sizes of the cloned genomic PC 5 UTR fragments were verified by PCR, using genomic DNA from four cows. Sequencing data confirms the existence of a 178 bp exon that contains the 68 and 110 bp sequence elements of the 5 UTR for PC mRNA. The 178 bp exon appears to be the first transcribed exon in PC and the 68 and 110 bp 5 UTR sequences are most likely generated by alternative transcription start sites. Genomic sequence data also confirms that the 3 end of the 89 bp element is a discrete 41 bp exon. Regions within the genomic sequence adjacent to the 178 and 41 bp exons of the PC 5 UTR contain binding sites for TBP, Sp1, Ap1 and/or CEBP transcription factors. These data provide information about the arrangement of exons in the 5 UTR of PC and about putative promoter regions.