Determination of late feathering genotype in Nagoya breed males using PCR-RFLP method
2009
Nakamura, A.(Aichi-ken. Agricultural Research Center, Nagakute (Japan)) | Kobayashi, M. | Noda, K. | Kondo, H. | Kansaku, N.
The avian endogenous virus gene (ev-21) is closely associated on the dominant sex-linked late feathering (LF) gene, K, on the Z chromosome of chickens. The LF and early feathering (EF) phenotypes are widely used for sex identification; when ksup(+)/ksup(+) sires are mated with K/- hens, male chicks are LF and females are EF. To introduce the feather sexing, two lines (EF male line and LF female line) are necessary. However, homozygous (K/K) and heterozygous (K/ksup(+)) late feathering genotypes exist in LF males and separating K/K and K/ksup(+) by phenotypic differences at hatch is not possible. Consequently, labor-intensive progeny testing has been required in order to establish LF female line. In White Leghorn, determination of the genotype can be conducted by using restriction fragment length polymorphism (RFLP) analysis of the K or ksup(+) gene flanking region segments amplified by the polymerase chain reaction (PCR). The present study was conducted to establish a specific PCR assay that distinguishes Nagoya breed K/K males from K/ksup(+) males. To investigate the differences of DNA sequence between K and ksup(+) genes in Nagoya breed, a total of 1456bp of the flanking regions in K and ksup(+) was sequenced. Comparison between K and ksup(+) in Nagoya breed, indicated that differences were detected at 7 positions. Transition was detected at position 213, 294 and 616. Transversion was detected at position 333, 694 and 794. Deletion was detected at position 189-193 in K. At position 292-295, ksup(+) contains a Mbo I site (GATC), whereas K did not contain the recognition site. Thus, these results indicate that the Mbo I RFLP can be used in Nagoya breed to differentiate the K and ksup(+) alleles.
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