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Development of Y‐chromosome‐Specific SCAR Markers Conserved in Taurine, Zebu and Bubaline Cattle
Author(s) -
Alves BCA,
Hossepian de Lima VFM,
MoreiraFilho CA
Publication year - 2010
Publication title -
reproduction in domestic animals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.546
H-Index - 66
eISSN - 1439-0531
pISSN - 0936-6768
DOI - 10.1111/j.1439-0531.2009.01491.x
Subject(s) - sexing , biology , zebu , rapd , bovine genome , genetics , polymerase chain reaction , genetic marker , genomic dna , inverse polymerase chain reaction , y chromosome , gene , microbiology and biotechnology , genome , genetic diversity , nested polymerase chain reaction , population , zoology , demography , sociology
Contents Sex pre‐selection of bovine offsprings has commercial relevance for cattle breeders and several methods have been used for embryo sex determination. Polymerase chain reaction (PCR) has proven to be a reliable procedure for accomplishing embryo sexing. To date, most of the PCR‐specific primers are derived from the few single‐copy Y‐chromosome‐specific gene sequences already identified in bovines. Their detection demands higher amounts of embryonic genomic material or a nested amplification reaction. In order to circumvent this, limitation we searched for new male‐specific sequences potentially useful in embryo sexing using random amplified polymorphic DNA (RAPD) analysis. Random amplified polymorphic DNA (RAPD) assay reproducibility problems can be overcome by its conversion into Sequence Characterized Amplified Region (SCAR) markers. In this work, we describe the identification of two bovine male‐specific markers (OPC16 323 and OPF10 1168 ) by means of RAPD. These markers were successfully converted into SCARs (OPC16 726 and OPF10 984 ) using two pairs of specific primers.Furthermore, inverse PCR (iPCR) methodology was successfully applied to elongate OPC16 323 marker in 159% (from 323 to 837 bp). Both markers are shown to be highly conserved (similarity ≥95%) among bovine zebu and taurine cattle; OPC16 323 is also highly similar to a bubaline Y‐chromosome‐specific sequence. The primers derived from the two Y‐chromosome‐specific conserved sequences described in this article showed 100% accuracy when used for identifying male and female bovine genomic DNA, thereby proving their potential usefulness for bovine embryo sexing.