Ahmadi Sadequllah, Takeda Misa, Ohkubo Takeshi
College of Agriculture, Ibaraki University, 3-21-1 Chuo, Ami, Ibaraki 300-0393, Japan.
Faculty of Animal Science, Afghanistan National Agricultural Sciences and Technology University (ANASTU), Kandahar, Afghanistan.
J Poult Sci. 2019 Oct 25;56(4):253-261. doi: 10.2141/jpsa.0180087.
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 () (24 bp indel), growth hormone () (T185G), and pituitary specific transcript factor 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 I and I endonucleases. Cloning followed by DNA sequencing was performed to ascertain the accuracy of the PCR-RFLP analysis for .Two alleles were found for the 24 bp indel, (T185G), and I, with the following respective allelic frequencies: 0.64 and 0.36, -T 0.91 and -G 0.09, and -A 0.64 and -B 0.36. Regarding the I polymorphism, three novel I recognition sites, as well as two reported I recognition sites, were detected in intron 5. Moreover, during sequence screening, two novel SNPs were found that generated restriction sites for I. Therefore, our results suggest that the indel, T185G, and I polymorphisms may be used as selection markers for Afghanistan Naked Neck chickens. Intron 5 of in the Afghani Naked Neck chicken was highly polymorphic compared to the reported gene (GenBank accession no. NC_006088.4).
我们研究了提高阿富汗本土裸颈鸡产量的方法。具体而言，我们分析了催乳素（PRL）基因（24 bp插入/缺失）、生长激素（GH）基因（T185G）和垂体特异性转录因子1（POU1F1）基因（内含子5）中的单核苷酸多态性（SNP）。从52只鸡采集血样并提取基因组DNA。使用I和I型内切酶，通过聚合酶链反应（PCR）、等位基因特异性PCR和PCR-限制性片段长度多态性（RFLP）分析上述位点的多态性。进行克隆后测序以确定PCR-RFLP分析对POU1F1的准确性。在PRL的24 bp插入/缺失、GH（T185G）和POU1F1中发现了两个等位基因，其各自的等位基因频率如下：PRL-插入0.64和PRL-缺失0.36，GH-T 0.91和GH-G 0.09，以及POU1F1-A 0.64和POU1F1-B 0.36。关于POU1F1多态性，在内含子5中检测到三个新的POU1F1识别位点以及两个已报道的POU1F1识别位点。此外，在序列筛选过程中，发现了两个产生POU1F1识别位点的新SNP。因此，我们的结果表明，PRL插入/缺失、GH T185G和POU1F1多态性可作为阿富汗裸颈鸡的选择标记。与已报道的POU1F1基因（GenBank登录号：NC_006088.4）相比，阿富汗裸颈鸡POU1F1的内含子5具有高度多态性。
