Submission Number: VAL-4-11-14

Abstract Number: 462

STRUCTURE OF THE GENE FOR UTEROFERRIN.

Jeffrey L Vallet* and Scott C Fahrenkrug

USDA, ARS, RLH US Meat Animal Research Center, Clay Center, NE ¹

Abstract:
PCR Amplification of porcine genomic DNA using primers from exon 2 of the gene for uteroferrin (UF; Simmen et al., 1989; DNA 8:543) resulted in a product containing two previously undescribed introns. To confirm the structure of the UF gene, a bacterial artificial chromosome (BAC) genomic library was screened using a uteroferrin specific cDNA probe. UF BAC clone DNA was digested with EcoRI resulting in a 14.5 kb fragment containing the gene for UF, which was subcloned into PBS II SK. Approximately 8 kb of DNA, including the entire gene for UF, was sequenced. The gene for UF spanned 2.5 kb and is comprised of 5 exons, a structure similar to the related tartrate resistant acid phosphatase genes of humans and rats, and different from the previously published UF gene. Comparison of the sequence of the region upstream of the UF gene transcription start site with a previously reported sequence (Gonzalez et al., 1994; DNA Cell Biol. 13: 365) revealed substantial differences. PCR analysis of genomic DNA was used to determine whether multiple UF genes were present in the pig genome. Primers from within the gene only amplified products consistent with the two newly discovered introns. Primers specific to the 5' region of the current UF gene sequence gave the appropriate product, while primers specific to the previously reported sequence did not. Genomic southern blotting with BamHI, StuI, MscI, SacI, DraI, BamHI + StuI, and BamHI + KpnI using a cDNA (bases 328-567; Simmen et al., 1989) as probe revealed single bands consistent with the current gene sequence. Thus, the previously reported gene structure and DNA sequence of the 5' promoter region may be incorrect, potentially deriving from cloning artifacts. These results necessitate a reexamination of the UF promoter region for potential transcription controlling elements.

Keywords: Swine, Uterus, Pregnancy

This abstract is being presented at: 8:00 AM in session:
Gene Regulation and Function II