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)用于肉鸡科鸟类性别鉴定的DNA探针的建立：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序列在禽类W染色体上具有广泛的保守性，利用该序列的引物进行PCR扩增，扩增出了许多龟科动物的雌性特异性条带。然而，在一些物种中，如东方白鹳（Ciconia boyciana），在Z染色体上也发现了EE0.6的同源序列。两个ee0.6相关序列；从白桦基因组DNA中克隆出W染色体上的XH0.6和Z染色体上的XH0.6 rsm，利用XH0.6和XH0.6 rsm引物扩增出W染色体上的雌性特异性条带和Z染色体上的雄性-雌性共同条带。(2)鸡的性染色体鉴定：从鸵鸟和鸸鹋的基因组dna中克隆出与鸡EE0.6（位于W染色体上）和鸡IREBP（位于Z染色体上）同源的ZOV3基因。这三个序列与鸡的同源序列保持了71 ~ 92%的一致性。荧光原位杂交（FISH）表明，这三个序列在鸵鸟、鸸鹋和食火鸡中都定位在一组特定的染色体上。然而，在雌性鸵鸟和雌性食火鸡中，其中一对的IREBP序列缺失。这些结果表明，Carinatae和Ratitae鸟类的性染色体都是从共同的祖先染色体对进化而来的，而鸵鸟和食火鸡的W染色体有少量分化。(3)在濒危鸟类交配程序中的应用：利用上述引物分别对C. boyciana的XH0.6和XH0.6 rsm进行PCR扩增，成功地扩增了濒危鸟类Nipponia的雌性特异性条带和雄性-雌性共同条带，并利用少量羽毛制备的基因组DNA进行了扩增。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