【摘要】 快慢羽作为Z染色体1对等位基因(K,k+)控制的性连锁表型,被广泛应用在配套系生产的雏鸡性别鉴定上。为明确慢羽鸡PRLR和SPEF2基因连接方式和融合基因双向转录特征,本试验通过半定量PCR扩增PRLR和SPEF2基因共有区域477bp片段,明确两基因在慢羽鸡的连接方式;以禽内源白血病病毒ev21和SPEF2基因的转录作为对照,通过特异引物反转录和S1酶切法确定融合基因的转录方向。试验明确了慢羽K基因中PRLR与dSPEF2、SPEF2与dPRLR的5’末端以"头碰头"方式连接,"头碰头"连接的477 bp共有区域仅PRLR发生转录;证明了dPRLR和dSPEF2的3’末端"尾对尾"融合区域为双向共转录区域;还发现S1酶切法验证基因转录方向存在局限性。该研究结果为深入挖掘鸡慢羽K基因的分子结构和调控机制奠定了基础。更多还原

【Abstract】 As a sex-linked phenotype controlled by a pair of Z chromosome alleles(K, k+), fast and slow feathering type has been widely used in the sex identification of chickens in mating lines. In order to clarify the connection pattern of PRLR and SPEF2 and the bidirectional transcription characteristics of the fusion gene in slow-feathering chickens, the semi-quantitative PCR was used to amplify the 477 bp fragment of the shared region of PRLR and SPEF2. The transcription direction of fusion gene was determined by specific primer reverse transcription, and S1 nuclease digestion of ev21 and SPFE2 was used as the control. It was confirmed that PRLR and dPRLR in K gene of chickens were reversely mapped on Z chromosome, and were connected with the 5’ terminal of dSPEF2 and SPEF2 in a “head-to-head” way, respectively. Only PRLR was transcribed in the common region of PRLR and SPEF2. It was proved that the “tail-to-tail” 3’ terminal of dPRLR and dSPEF2 was a bidirectional co-transcription region. In addition, S1 nuclease digestion method was found to have limitation in verifying the direction of gene transcription. The results lay a foundation for further exploring the molecular structure and regulatory mechanism of K gene in slow feathering chicken.更多还原