中国地方家鸭ZW性染色体上CHD1基因序列差异的检测新方法

A New Detecting Method for the Sequence Difference of CHD1 Gene on the Z and W Sex Chromosomes in Chinese Native Domestic Duck Breeds

旨在发掘一种能高效和快速检测鸭ZW性染色体上CHD1基因序列差异的方法,以解决鸭产业中胚胎期无损伤性别鉴定、胚胎期性别人工鉴定误判和新生期翻肛鉴别损伤等技术问题。本研究利用鸟类CHD1基因序列的DNA扩增片段长度多态性特点,成功获得1对能够在中国地方家鸭品种中广泛使用的性别鉴定引物HPF/HPR,对所获得的PCR产物进行了克隆、测序和比对,并通过实例对该性别分子鉴定方法的有效性和准确性进行了验证。结果显示,经2%琼脂糖凝胶电泳就能清晰区分由引物HPF/HPR获得大小为495bp的CHD1-Z和351bp的CHD1-W PCR产物,并且雌性(ZW型)同时出现明显的双条带,而雄性(ZZ型)则出现单一条带。经证实,本研究提供的检测中国地方家鸭ZW性染色体上CHD1基因序列差异的方法直观可靠,同时也为中国地方家鸭的性别分子生物学鉴定提供了一个高效准确的分子遗传标记。

In the current study, a new method for detecting the sequence difference of duck Chromo-helicase-DNA-Binding 1 （CHD1） genes on Z and W sex chromosomes was established through amplifying DNA fragment length polymorphism, which aimed to solve some technical problems, such as the sex identification with nondamage, the misclassific rate of artificial sex identification during embryonic period and the injury coming from the rectal identification during neonatal period. In this study, an improved gender identification method was developed by DNA amplified fragment length ploymorphism between CHD1-Z and CHD1-W genes of birds. The gender identification PCR primers HPF/HPR were used, and the PCR product was cloned, sequenced and blasted; then the effectiveness and accuracy of this method had been explored by examples. 2% agarose gels were used to easily distinguish the 495 bp CHD-Z and the 351 bp CHD-W PCR amplicons, and the female （ZW） displayed two visible bands, whereas only a single band was found in the male （ZZ）. The results indicate that the new detecting method for detecting the sequence difference of duck CHD1 genes is visual and reliable, and the molecular marker provided by this study is highly efficient and precise in the sex molecular biological identification of Chinese native domestic duck breeds.