Determination of Polymorphisms in Pituitary Genes of the Native Afghani Naked Neck Chicken
Determination of Polymorphisms in Pituitary Genes of the Native Afghani Naked Neck Chicken
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DOI：
10.2141/jpsa.0180087
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发表时间：
2019-10
期刊：
影响因子：
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通讯作者：
Sadequllah Ahmadi;Misa Takeda;T. Ohkubo
中科院分区：
文献类型：
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作者：
Sadequllah Ahmadi;Misa Takeda;T. Ohkubo
我们研究了提高阿富汗本土裸颈鸡产量的方法。具体来说，我们分析了催乳素 (PRL)（24 bp 插入缺失）、生长激素 (GH) (T185G) 和垂体特异性转录因子 1 (PIT-1)（内含子 5）基因中的单核苷酸多态性 (SNP)。从 52 只鸟身上采集血液样本并提取基因组 DNA。使用 TaqI 和 MspI 核酸内切酶通过 PCR、等位基因特异性 PCR 和 PCR 限制性片段长度多态性 (RFLP) 分析上述基因座的多态性。进行克隆后进行 DNA 测序，以确定 PIT-1 PCR-RFLP 分析的准确性。发现 PRL 24 bp 插入缺失、GH (T185G) 和 PIT-1/TaqI 的两个等位基因，其等位基因频率如下：PRL-In 0.64 和 PRL-Del 0.36、GH-T 0.91 和 GH-G 0.09，以及PIT-1-A 0.64 和 PIT-1-B 0.36。关于PIT-1/MspI多态性，在内含子5中检测到三个新的MspI识别位点以及两个报道的MspI识别位点。此外，在序列筛选过程中，发现了两个新的SNP，它们产生了MseI的限制性位点。因此，我们的结果表明，PRL indel、GH T185G 和 PIT-1/TaqI 多态性可用作阿富汗裸颈鸡的选择标记。与报道的原鸡 PIT-1 基因（GenBank 登录号 NC_006088.4）相比，阿富汗裸颈鸡 PIT-1 的内含子 5 具有高度多态性。
We investigated means to improve the production of the indigenous Naked Neck chicken in Afghanistan. Specifically, we analyzed single nucleotide polymorphisms (SNPs) in the prolactin (PRL) (24 bp indel), growth hormone (GH) (T185G), and pituitary specific transcript factor 1 (PIT-1) (intron 5) genes. Blood samples were collected from 52 birds and genomic DNA was extracted. Polymorphisms in the mentioned loci were analyzed by PCR, allele-specific PCR, and PCR-restriction fragment length polymorphism (RFLP) using TaqI and MspI endonucleases. Cloning followed by DNA sequencing was performed to ascertain the accuracy of the PCR-RFLP analysis for PIT-1.Two alleles were found for the PRL 24 bp indel, GH (T185G), and PIT-1/TaqI, with the following respective allelic frequencies: PRL-In 0.64 and PRL-Del 0.36, GH-T 0.91 and GH-G 0.09, and PIT-1-A 0.64 and PIT-1-B 0.36. Regarding the PIT-1/MspI polymorphism, three novel MspI recognition sites, as well as two reported MspI recognition sites, were detected in intron 5. Moreover, during sequence screening, two novel SNPs were found that generated restriction sites for MseI. Therefore, our results suggest that the PRL indel, GH T185G, and PIT-1/TaqI polymorphisms may be used as selection markers for Afghanistan Naked Neck chickens. Intron 5 of PIT-1 in the Afghani Naked Neck chicken was highly polymorphic compared to the reported Gallus gallus PIT-1 gene (GenBank accession no. NC_006088.4).