Investigation of PRL-RsaI and HaeIII gene polymorphisms in Anatolian water buffaloes bred by using PCR-RFLP method

ABSTRACT The objective of this study was to investigate polymorphisms both in exons 1 and 3 of prolactin (PRL) gene for milk productivity of Anatolian water buffalo breed in Sivas province in Turkey. Blood samples were collected from 129 male and female water buffaloes and DNA was isolated by using phenol/chloroform method. Samples of DNA were amplified and resulting PCR products were digested with RsaI (for exons 1 and 3) and HaeIII (for exon 1). Allelic polymorphisms were determined by separation of fragments obtained from digested PCR products in 3% agarose gel electrophoresis. AA genotype (HaeIII) and BB genotype (RsaI) of exon 1 and only AA genotype (RsaI) of exon 3 were obtained. No polymorphisms were determined in Anatolian water buffalo breed and all loci were found as monomorphic. It can be stated that Anatolian water buffalo has a higher milk and milk fat yields since BB genotype was obtained.摘要 本研究旨在探究土耳其锡瓦斯省安纳托利亚水牛催乳素（prolactin, PRL）基因第1外显子与第3外显子的多态性及其对泌乳性能的影响。 本研究采集了129头公、母安纳托利亚水牛的血液样本，采用酚-氯仿法提取基因组DNA。 对DNA样本进行聚合酶链式反应（Polymerase Chain Reaction, PCR）扩增，所得PCR产物分别用限制性内切酶RsaI（针对第1、3外显子）与HaeIII（针对第1外显子）进行酶切。 通过3%琼脂糖凝胶电泳分离酶切后的PCR产物片段，以此鉴定等位基因多态性。 结果显示，第1外显子经HaeIII酶切后得到AA基因型、经RsaI酶切后得到BB基因型，第3外显子经RsaI酶切后仅得到AA基因型。 本研究未在安纳托利亚水牛群体中检测到多态性，所有位点均为单态性位点。 结合本次检测到的BB基因型可推测，安纳托利亚水牛具备较高的泌乳量及乳脂产量。