Present address: Applied Biosystems, 850 Lincoln Centre Dr., Foster City, CA 94404, USA.
Interfamilial characterization of a region of the ZFX and ZFY genes facilitates sex determination in cetaceans and other mammals
Article first published online: 1 JUL 2005
Volume 14, Issue 10, pages 3275–3286, September 2005
How to Cite
MORIN, P. A., NESTLER, A., RUBIO-CISNEROS, N. T., ROBERTSON, K. M. and MESNICK, S. L. (2005), Interfamilial characterization of a region of the ZFX and ZFY genes facilitates sex determination in cetaceans and other mammals. Molecular Ecology, 14: 3275–3286. doi: 10.1111/j.1365-294X.2005.02651.x
- Issue published online: 1 JUL 2005
- Article first published online: 1 JUL 2005
- Received 21 February 2005;revisionreceived 26 April 2005;accepted 20 May 2005
- 5’ exonuclease assay;
- sex determination;
Sequence polymorphism of homologues ZFX and ZFY, in a 604-base pair exon region, was examined in 10 known males and 10 known females across seven cetacean families and used to design a simple, highly sensitive and widely applicable fluorescent 5′ exonuclease assay for gender determination in cetaceans. Multiplex amplification, cloning, and sequencing of these previously uncharacterized regions revealed (i) eight fixed differences between ZFX and ZFY, (ii) 29 variable sites between ZFX and ZFY and (iii) very low interspecific nucleotide diversity for both ZFX and ZFY across all families examined. We developed a 5′ exonuclease assay that produces a small (105 bp) polymerase chain reaction (PCR) product from both the X and the Y chromosome orthologs, and used double-labelled fluorescent probes to distinguish between the two genes in a real-time PCR assay that is highly reproducible and sensitive. We demonstrated sex specificity for 33 cetacean species in nine families. Given the availability of conserved primers and sequence information for many mammalian species, this approach to designing sexing assays for a wide range of species is both practical and efficient.