采用血液、羽毛等材料鉴别鸡性别PCR方法的应用研究

Sex Identification of Chicken with Blood, Feathers and Other Materials by PCR

公母鸡染色体上的染色体螺旋蛋白基因(chromobox-he-licase-DNA binding gene,CHD 1基因)的内含子基因序列存在差异,通过PCR方法可扩增该基因内含子的保守序列,可对鸡等非平胸禽类的性别进行鉴定。本研究首先采集42只28日龄肉鸡抗凝血,完成血液DNA PCR鉴定性别,结果表明,42只商品肉鸡中22只公鸡、20只母鸡,与剖检结果相同;其次,采集20只90日龄SPF鸡(13只公鸡、7只母鸡)和15只2日龄SPF鸡的抗凝血和羽毛,完成其DNA的PCR鉴定以及与微量血的PCR检测结果的比较。结果表明,90日龄、2日龄SPF鸡的微量血PCR与抗凝血、羽毛提取DNA的PCR鉴定结果均相同,其中,90日龄SPF鸡与其外观性别完全吻合;另外,采集12枚15日龄鸡胚的尿囊液、羽毛、肌肉,提取DNA后PCR方法可扩增出区分性别的特异性条带,鉴定结果一致,12枚15日龄鸡胚中均为7个雄性、5个雌性。本研究表明,微量血PCR和血液、羽毛等材料提取DNA后的PCR方法均可准确快速鉴定鸡和鸡胚的性别,可作为家禽育种过程中常规鉴定方法的有力补充,适合大型养殖场规模化鉴定,也满足科研领域区分鸡和鸡胚性别的需求。

The intron gene sequence of CHD1 gene（chromobox-he-licase-DNA binding gene） was different between the chromosomes of hens and roosters.PCR（Polymerase Chain Reaction） was used to amplified the conserved sequence of the intron gene sequence in order to identified the sex of non-ratite birds such as chickens. First of all,we collected 42 anticoagulant blood samples of 28-day-old broilers,extracted DNA from the blood and used PCR to indentify the sex.The results showed that there were 22 roosters and 20 hens^the same as the necropsy result. Secondly,we collected anticoagulant blood and feathers samples of 20 90-day-old SPF chickens（13 roosters, 7 hens） and 15 2-day-old SPF chickens, DNA was extracted and was used in PCR. Meanwhile,PCR amplification were taken on microscale blood samples and the result were identical to the PCR（with DNA） result. Among the result ,all the 90-day-old SPF chickens＂ sex were identical to their appearance sex. Thirdly, we collected feathers, muscles and allantoic fluid samples of 12 15-day-old chicken embrysos and then extracted DNA. Sex- specific bands were amplified by PCR and the result showed that there were 7 male embrysos and 5 female embrysos. In conclusion, both PCR with microscale blood samples and PCR with DNA extracted from blood and feathers could quickly and accurately identify the sex of embrysos and chickens. It could be a strong complement to conventional methods of sex identification among poultry breeding process.