Submitted to: Society for the Study of Reproduction Annual Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 4/24/2006
Publication Date: 7/20/2006
Citation: Kim, J.G., Ford, J.J., Rohrer, G.A., Nonneman, D.J. 2006. Molecular cloning of porcine OGT cDNA and mapping to the X chromosome [abstract]. Biology of Reproduction. Supplement:154.
Technical Abstract: O-linked N-acetylglucosamine transferase (OGT) is located on the human X chromosome and catalyzes the post-translational addition of a single N-acetylglucosamine in O-glycosidic linkage to serine or threonine residues of proteins. N-acetylglucosamine glycosylation is implicated in nuclear transport of proteins, fertilization, and regulation of the cell cycle and is essential for embryogenesis. Therefore, we hypothesized that OGT may play a role in swine reproductive physiology. We isolated a pig OGT cDNA, by screening the MARC 2PIG EST library with primers designed from partial EST sequences. A partial OGT cDNA containing 4623 bp was identified and it did not contain the 5’ untranslated region (UTR) or the translation start site. To obtain a full-length sequence, a forward primer was designed from an EST sequence in GenBank and a reverse primer was designed from the partial OGT cDNA from this study. Using 2PIG cDNA as template, a 1266 bp cDNA fragment was amplified by PCR and subcloned. The fragment contained the 5’ UTR and part of the coding region. The composite cDNA contained 5391 bp (90.6% identical with human) encoding 1046 amino acids, which was 99.8% identical to the human sequence (GenBank accession DQ400859). In order to map OGT using single nucleotide polymorphisms (SNP), we amplified and sequenced a region containing the putative intron 16 of the OGT gene. Interestingly, amplification and sequencing of genomic DNA of the reference population indicated a length polymorphism in intron 16 of the OGT gene between Chinese breeds and White composite pigs. The size of this intron in Minzu, Meishan and Fengjing crossbred boars was 222 bp, while the size in White composite pigs and Duroc was 510 bp due to an inserted 282 bp PRE-1 SINE element. Using this pattern of polymorphism in the reference family, the OGT gene was mapped to 67 cM on the porcine X chromosome. Amplification of genomic DNA of one-quarter Meishan boars indicated that this polymorphism can determine the breed of inheritance of this region in Chinese and European crossbreds. This length polymorphism will be useful in further defining the FSH and testis size QTL located on the porcine X chromosome.