Molecular characterization and cytological mapping of a non-repetitive DNA sequence region from the W chromosome of chicken and its use as a universal probe for sexing Carinatae birds
Molecular characterization and cytological mapping of a non-repetitive DNA sequence region from the W chromosome of chicken and its use as a universal probe for sexing Carinatae birds
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DOI：
10.1023/a:1018461906913
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发表时间：
1997-04-01
影响因子：
2.6
通讯作者：
Mizuno, S
中科院分区：
文献类型：
--
作者：
Ogawa, A;Solovei, I;Mizuno, S
利用鸡单W染色体基因组文库，从鸡W染色体上克隆出约25 kb的非重复基因组DNA区域。利用有丝分裂和灯刷染色体的荧光原位杂交（FISH）技术将该区域定位在W染色体上的EcoRI家族和XhoI家族之间。从该区域克隆的0.6 kb EcoRI片段（EE0.6）由外显子捕获程序和侧翼序列组成。对隶属于8个不同分类目的18种鸟类的雄性和雌性基因组dna进行Southern blot杂交，发现在Carinatae鸟类的W染色体上广泛保守有与EE0.6非常相似的序列。这些鸟的雌性可以通过一个明确的雌性特定波段的存在来确定。对于许多鸟类，也可以利用鸡EE0.6侧翼序列的一组引物，通过聚合酶链反应（PCR）来确定雌性。
A non-repetitive genomic DNA region of about 25 kb was cloned from the W chromosome of chicken using a genomic library prepared from a single W chromosome of the chicken. This region was mapped by fluorescence in situ hybridization (FISH) with mitotic and lampbrush chromosomes to a position between the major EcoRI family and the pericentromeric XhoI family on the W chromosome. A 0.6-kb EcoRI fragment (EE0.6) subcloned from this region consists of a sequence that can be obtained by the exon-trapping procedure and flanking sequences. Sequences, which are closely similar to that of EE0.6, are widely conserved on the W chromosomes of Carinatae birds, as revealed by Southern blot hybridization to HindIII-digested female and male genomic DNAs from 18 species of birds belonging to eight different taxonomic orders. The female sex of those birds can be determined by the presence of an unambiguous female-specific band. For many species of birds, the female sex can also be determined by polymerase chain reaction (PCR) using a set of primers from the flanking sequences in the chicken EE0.6.