Novel Polymorphisms of ZRSR2 and GPM6B Gene Homologs and Their Use in Sex Identification of Bovine and Porcine Species

Accurate and cost-effective PCR based sex identification is important in animal production because it gives producers the ability to determine the sex of embryos prior to transfer, saving time and money. The most efficient PCR sex identification assays work by using a single primer pair to amplify a specific target region located on the Y-chromosome and a second, separate target region on the X-chromosome. This thesis reports the design of two novel assays. The first assay was designed to target the Zinc finger CCCH-type, RNA binding motif and serine/arginine rich 2 (ZRSR2) gene found on the X-chromosome and its Y-chromosome homolog, ZRSR2Y, in European domestic cattle (Bos taurus). It creates X- and Y-specific amplicons of 149 bp and 116 bp long respectively and is effective for sex identification of Bos taurus, African domestic cattle (Bos indicus), bison (Bison bison), domestic goat (Capra hircus), domestic sheep (Ovis aries), and domestic yak (Bos grunniens). The second assay was designed to target the glycoprotein M6B (GPM6B) gene found on the X-chromosome in domestic pigs (Sus scrofa) and a homologous pseudogene on the Y-chromosome. It creates X- and Y-specific amplicons of 244 bp and 201 bp long, respectively. The presence of a 16 bp intron deletion variant (rs1111696537) on the X-specific amplicon in a subset of the population was confirmed. Despite the variant, the primer pair is effective for sex identification of domestic pigs