Use of the TSPY gene for sexing cattle

Abstract

The Y-encoded, testis-specific protein (TSPY) is a Y-specific gene. The copy numbers of TSPY range from 20 to 60 in men and up to 200 in bulls. In this study, we examined the possibility of using the TSPY gene to sex cattle. DNA from blood samples of 100 Nelore cattle (50 males and 50 females) from the Nelore Cattle Breeding Program (PMGRN) was screened for TSPY by PCR using TSPY-specific primers. The assay was highly specific since all male samples were TSPY-positive and all female samples were negative. Positive results were also obtained at low DNA concentrations (less than 1 ρg/µL). These results showed that TSPY was a good male-specific marker, the usefulness of which was enhanced by the high copy number of the gene. This is the first report to demonstrate the applicability of TSPY for sexing cattle.

Sexing single bovine blastomeres using TSPY gene amplification

“…In a previous study, we reported that PCR using primers derived from a TSPY sequence allows the detection of the Y chromosome in male animals (Nelore cattle) even at low DNA concentrations. These preliminary results showed the potential of this gene as a male-specific marker (Lemos et al, 2005).…”

“…The application of polymerase chain reaction (PCR) for the analysis of a single cell presents a challenge since limited DNA is available in one single diploid cell, and this can lead to a reduction in amplification efficiency. Lemos et al (2005) described a study on the TSPY (testis-specific protein Y-encoded) gene as a male-specific marker for cattle sexing in very low DNA concentrations. The TSPY gene is advantageous compared to other markers because it is a Y-specific gene of multiple copies both in human and bovine genomes.…”

“…The TSPY gene is advantageous compared to other markers because it is a Y-specific gene of multiple copies both in human and bovine genomes. After embryo biopsy, the DNA from only one cell is amplified, so, using genes with multiple copies, we could increase the efficiency of the amplification (Bartmann et al, 2004;Lemos et al, 2005).…”

“…In the present study, we tested a nested PCR-based protocol, modified from Lemos et al (2005), in order to sex single cells from bovine preimplantation embryos using a sequence of the TSPY gene (also known as DYS14 in humans).…”

Amplificación del gen TSPY1 como método de sexaje en bovinos

“…Por otro lado, la detección del sexo a partir de una muestra de ADN mediante PCR puede ser realizada usando primers específicos de distintos genes como el SRY, TSPY1, y otros. Dado que el gen TSPY1 es multicopia, es decir, presenta varias copias en bajas concentraciones de ADN, resulta más útil y eficiente como marcador masculino a diferencia de otros genes como el SRY que es de 1 sola copia (Lemos et al, 2005).…”