Development of DNA probes for identification of sexes of a wide range of avian species and their application to the mating program of endangered species

(1)鸡性别鉴定用DNA探针的研制：从鸡W染色体上克隆了一个非重复基因组DNA片段EE0.6。EE0.6序列被证明是广泛保守的鸟类W染色体上，和PCR使用一套引物从这个序列扩增的女性特异性带在一些Carinerus物种。然而，在某些物种中，如东方白色鹳（Ciconia boyciana），Z染色体上也发现了与EE 0.6同源的序列。从中国对虾中克隆了两个与EE 0.6相关的序列：位于W染色体上的XH 0.6和位于Z染色体上的XH 0.6RSM。Boyciana的基因组DNA，用引物XH0.6和XH0.6RSM扩增出W染色体上的雌性特异带和Z染色体上的雌雄通用带。男孩。(2)鸡亚科鸟类性染色体的鉴定：与鸡EE0.6（W染色体上）和鸡IREBP和 关于我们 从鸵鸟和鸸鹋的基因组DNA中克隆了ZOV 3基因（均位于Z染色体上）。这三个序列的Ratitae物种保持71-92%的同源性，鸡的对应物。荧光原位杂交（FISH）使用这些克隆的序列从Ratitae，表明所有这三个序列定位在一组特定的染色体在鸵鸟，鸸鹋和食火鸡。然而，雌鸵鸟和雌食火鸡中的一对IREBP序列缺失。这些结果表明，隆头鸟和灰雀科鸟类的性染色体都是由共同的祖先染色体对进化而来的，鸵鸟和食火鸡的W染色体略有分化。(3)在濒危鸟类交配程序中的应用：用上述引物XH0.6和XH0.6RSM扩增出的雌特异带和雌雄共带。boyciana，也分别成功地为日本，一种濒危物种的lbis，使用基因组DNA制备的一些羽毛。少

(1) Development of DNA probes for identification of sexes of Carinatae species of birds : A non-repetitive genomic DNA fragment, EE0.6, from the chicken W chromosome was clones. The EE0.6 sequence was shown to be conserved widely on the avian W chromosome, and PCR using a set of primers from this sequence amplified a female-specific band in a number of Carinatae species. However, in some species, like Oriental white stork (Ciconia boyciana), a homologous sequence to EE0.6 was also found on the Z chromosome. The two EE0.6-related sequences ; XH0.6 on the W chromosome and XH0.6RSM on the Z chromosome, were cloned from C. boyciana, and PCR using sets of primers from XH0.6 and XH0.6RSM amplified a female-specific band form the W chromosome and male-female-common bands from the Z chromosome from the genomic DNA preparations of C. boyciana.(2) Identification of sex chromosomes of Ratitae birds : Homologous genomic sequences to the chicken EE0.6 (on the W chromosome) and the chicken IREBP and … More ZOV3 genes (both on the Z chromosome) were cloned from genomic DNAs of ostrich and emu. Those three sequences from the Ratitae species maintained 71-92% identities to those of the chicken counterparts. Fluorescence in situ hybridization (FISH) using these cloned sequences from the Ratitae, demonstrated that all the three sequences were localized on a particular set of chromosomes in ostrich, emu and cassowary. However, the IREBP sequence was missing from one of the pairs in the female ostrich and the female cassowary. These results demonstrated that sex chromosomes of both Carinatae and Ratitae birds were evolved from the common ancestral pairs of chromosomes and that a little differentiation of W chromosome was evident in the ostrich and the cassowary.(3) Application to the mating program of endangered avian species : PCR amplification of the female-specific and the male-female-common bands using the sets of primers from the above XH0.6 and XH0.6RSM of C. boyciana, respectively, was also successful for Nipponia, an endangered species of lbis, using the genomic DNA prepared from a few feathers. Less