Search for the specific gene region in Silkie chicken

乌骨鸡是我国传统医学中的重要品种，但与乌骨鸡黑色素形成和沉积相关的基因尚不清楚。采用PCR差减法构建了丝羽乌骨鸡和白色来航鸡的基因组差减文库。我们对这些文库中的178个克隆进行了测序，发现了86个独特的克隆。对丝羽乌骨鸡和白色来航鸡进行了PCR-RFLP检测。在13对引物中，丝羽乌骨鸡和白色来航鸡之间存在多态性。用两对PCR-RFLP引物检测了53只丝羽乌骨鸡、19只日本土鸡和6只商品鸡催乳素基因启动子区24 bp的插入片段，结果表明，丝羽乌骨鸡催乳素基因启动子区24 bp的插入片段与其它品种的等位基因频率存在5%的显著差异。46只乌骨鸡为Del/Del基因型，7只乌骨鸡为In/Del基因型，其余均为In/Del基因型 ...更多信息 乌骨鸡无In/In基因型。而其它鸡群中分别有11只、8只和6只鸡出现Del/Del、In/Del和In/In基因型。2 × 3卡方检验表明，丝羽乌骨鸡与其它鸡种的基因型频率差异极显著（P<0.01）。这些结果表明，丝羽乌骨鸡催乳素基因启动子区24 bp插入片段的基因型频率与其它鸡有显著差异。丝羽乌骨鸡（Gallus gallus）在我国传统医药中占有重要地位，但其产蛋率很低，多有就巢现象。催乳素基因被认为是母鸡就巢的启动和维持所必需的。与红林鸡相比，白色来航鸡催乳素基因启动子区存在24 bp的插入片段，该片段的缺失将维持催乳素基因启动子的正常功能。在本研究中，24-bp的插入基因型在第四代的乌骨鸡选择用于产蛋的群体和插入位点的基因型和生产性能之间的关系进行了研究。从大分县农林水产研究中心家畜研究所选择的第4代丝羽乌骨鸡群中采集69只雄性丝羽乌骨鸡和242只雌性丝羽乌骨鸡的血液样品。从血样中提取基因组DNA，用PCR扩增催乳素基因启动子片段。将PCR产物在3.0%琼脂糖凝胶上电泳以进行基因分型。本研究共发现Del/Del丝羽乌骨鸡216只，In/Del丝羽乌骨鸡26只，In/In丝羽乌骨鸡未发现。方差分析表明，插入位点的基因型与生产性能相关，但与生长性能和产蛋量无关。In/Del乌骨鸡151 - 450日龄产蛋率的分布峰值高于De/Del乌骨鸡。少

Silkie chicken (Gallus gallus) have played an important role in traditional Chinese Medicine, however the genes associated with the specific appearance and deposition of the melanin pigment are unknown. The three genomic subtraction libraries were constructed using PCR subtraction method between a Silkie chicken and a White Leghorn chicken. We sequenced 178 clones of these libraries and found the 86 unique clones. The primer pairs for PCR-RFLP system were examined for the Silkie chicken and the White Leghorn chicken. The polymorphism between Silkie chicken and White Leghorn was found in the thirteen primer pairs. The 5% level significant differences of allele frequency between Silkie chicken and the other breeds were found in two PCR-RFLP primer pairs.The 24-bp insertion of the promoter region of prolactin gene was studied for 53 Silkie fowls, 19 Japanese native chickens and 6 commercial chickens. Forty-six Silkie fowls have genotype Del/Del, 7 Silkie fowls have genotype In/Del, howeve … More r no Silkie fowls have genotype In/In. On the other hand, 11, 8 and 6 birds of other chickens showed genotypes Del/Del, In/Del and In/In, respectively. The chi-squared test using 2x3 table showed the significant difference (P<0.01) of the genotype frequencies between Silkie fowls and other chickens. These results show the significant difference of the genotype frequencies for the 24-bp insertion of the promoter region of prolactin gene between Silkie fowls and other chickens.Silkie chicken (Gallus gallus) have played an important role in traditional Chinese Medicine, however broodiness is often observed and egg production ratio is very small in Silkie chicken. It is thought that prolactin gene is an essential for the start and maintain of broodiness in hens. The 24-bp insertion is found in the promoter region of prolactin gene of White Leghorn as compare with Red Jungle fowl and the absence of the 24-bp insertion would maintain normal function of the prolactin promoter. In this study, the 24-bp insertion was genotyped in the fourth generation of the Silkie population selected for egg production and the relationship between the genotype of the insertion locus and performance traits was investigated. Blood samples of 69 male Silkies and 242 female Silkies were collected form the fourth generation of Silkie population selected for egg production in Livestock Research Institute of Oita Prefectural Agriculture, Forestry and Fisheries Research Center. Genomic DNA was isolated from blood samples and the fragment of the promoter of prolactin gene was amplified using PCR. The PCR products were electrophoresed on 3.0% agarose gels for genotyping. The 216 Del/Del Silkies, 26 In/Del Silkies and no In/In Silkie were found in this study. The relationship between the genotype of the insertion locus and the performance traits was analyzed by analysis of variance, however there was no relationship for growth traits and egg production. The In/Del Silkies have higher peak of the distribution of 151d-450d egg production ratio than the De/Del Silkies. Less