QDPR基因克隆及其在乌蒙凤鸡不同生长阶段组织表达研究

旨在克隆乌蒙凤鸡醌型二氢生物喋呤还原酶（quinoid dihydropteridine reductase，QDPR）基因，并检测其在不同性别乌蒙凤鸡不同生长阶段各组织中的表达差异，以研究QDPR基因在乌蒙凤鸡生长发育过程中的调控作用。本试验对乌蒙凤鸡QDPR基因CDS区进行PCR扩增和克隆，并对得到的序列进行生物信息学分析，通过实时荧光定量PCR检测QDPR基因在4、12、20和28周龄乌蒙凤鸡公鸡及母鸡的心、肝、脾、肺、肾、胸肌和腿肌组织中的相对表达量。结果表明，QDPR基因开放阅读框长度为417 bp，共编码139个氨基酸，编码蛋白为酸性不稳定蛋白，乌蒙凤鸡的QDPR基因序列与原鸡、日本鹌鹑和珠鸡的QDPR基因同源性较高；QDPR基因在不同性别乌蒙凤鸡不同生长阶段各组织中均有表达，其中，公鸡4、12和20周龄肺组织中QDPR基因表达量最高，极显著高于同一周龄其他组织（P<0.01），母鸡4周龄肝、脾、肺中QDPR基因相对表达量为所有时期最高，极显著高于12、20周龄（P<0.01），公鸡大多数组织中QDPR基因表达量随时间增长总体呈现出先升后降的趋势，母鸡为先降后升。结果显示，QDPR基因在乌蒙凤鸡内脏组织中的表达均高于肌肉组织，且公母鸡不同时期表达情况有所差异，试验结果可为进一步研究QDPR基因在鸡生长发育中的调控作用提供参考。

Cloning of QDPR Gene and Its Expression in Various Tissues in Different Growth Stages of Wumeng Crested Chicken

The aim of this study was to clone quinoid dihydropteridine reductase (QDPR) gene and detect its expression in different tissues, different growth stages of Wumeng Crested chicken of different genders, so as to study the regulatory role of QDPR gene in the growth and development of Wumeng Crested chicken. In this experiment, the CDS region of QDPR gene of Wumeng Crested chicken was amplified and cloned by PCR, bioinformatics analysis of the sequences was carried out, and real-time fluorescent quantitative PCR analysis was performed to detect the expression of QDPR gene in heart, liver, spleen, lung, kidney, chest muscle and leg muscle of Wumeng Crested chicken in 4, 12, 20, and 28 weeks old. The results showed that the length of the open reading frame of QDPR gene was 417 bp, and 139 amino acids were encoded in total. The encoding protein was acidic and unstable. The QDPR gene sequence of Wumeng Crested chicken was highly homologous with the Gallus gallus, Coturnix japonica and Numida meleagris; QDPR gene was expressed in all tissues of Wumeng Crested chickens of different genders at different stages, the relative expression level of QDPR gene was the highest in the lung tissue of cock in 4,12 and 20 weeks old, which was extremely significantly higher than that in other tissues at the same age (P<0.01). The relative expressions of QDPR gene in liver, spleen and lung of 4-week-old hens were the highest in all stages of growth, which was extremely significantly higher than those in the stages of 12- and 20-week-old (P<0.01). The relative expressions of QDPR gene in the most of tissues of cock generally showed a trend of increasing first and then decreasing with time, while decreasing first and then increasing in hens. The results indicated that the expressions of QDPR gene in visceral tissues of Wumeng Crested chickens were higher than that in muscle tissues, and the expression in cock and hen were different in different growth stages. The experimental results can provide a reference for further research on the regulation mechanism of QDPR gene in chicken growth and development.