【摘要】 本试验旨在探讨鸡羽速基因型Hae Ⅲ酶切鉴定方法的分子基础。以羽型明确的6个品系(太行鸡、坝上长尾鸡、大午粉鸡、海兰灰鸡、海兰褐鸡、海兰灰祖代四系鸡)313份鸡基因组为模板,利用Hae Ⅲ酶切的RFLP试验进行羽速验证,并对ev21占位区(OS)和非占位区(US)共有序列以及OS区特有序列进行酶切试验。结果表明:1)Blast分析发现1 450bp为鸡ev21的US区域片段。海兰灰及其祖代四系和大午粉祖代的羽速基因型与HaeⅢ酶切结果完全一致,但太行慢羽公鸡、慢羽母鸡、坝上长尾慢羽公鸡和海兰褐慢羽鸡的一致率分别为40.0%、27.6%、28.6%和0.0%;2)太行鸡和坝上长尾鸡ev21的OS和US区共有序列538bp酶切鉴定结果与表型一致率达到92%以上,其他品系(除海兰褐快羽公鸡为0.0%)均为100.0%;3)鸡ev21的OS区特异序列1 440bp的扩增阳性率在太行慢羽公鸡、慢羽母鸡和快羽公鸡中的阳性率分别为94.1%、65.5%和0.0%,在海兰灰祖代鸡中分别为100.0%和0.0%,并且1 440片段不能被Hae Ⅲ切开。综合分析6个品系ev21的OS和US区结构特征,认为US区Hae Ⅲ酶切位点变异不能作为慢羽和内源病毒ev21的鉴定依据,而OS区的ev21插入与1 440bp序列Hae Ⅲ酶切位点的A→G突变和八碱基重复是紧密连锁的。更多还原

【Abstract】 The aim of this study was to explore the molecular basis of Hae Ⅲ digestion detection of genotypes of chicken feather speed in chicken.Three hundred and thirteen individuals in 6strains of chickens(Taihang,Bashang Long-tail,Dawufen,Hy-line Gray,Hy-line Brown,progenitor of Hy-line Gray)with definite phenotype of feather speed were detected with RFLP.The common region of occupied site(OS)and unoccupied site(US)for 538 bp,also the specific region of OS for1 440 bp,were digested with HaeⅢ.The results showed that:1)The fragment for 1 450 bp was the unoccupied-site repeated sequence for ev21 ascertained by Blast.The digestion detection results of HaeⅢ for 1 450 bp were completely consistent with the phenotype of HY-line Gray and progenitors of HY-line Gray and Dawufen,but their consistency with Taihang slow-feather cock,Taihang slow-feather hen,Bashang Long-tail slow-feather cock and HY-line Brown slow-feather chicken were 40.0%,27.6%,28.6%and 0.0%,respectively.2)The consistency was above 92% between digestion results of 538 bp and phenotype in Taihang and Bashang Long-tail chicken,and the other strains were 100.0%except for HY-line Brown fast-feather cock with 0.0%.3)The positive rate of 1 440 bp fragment PCR in slow-feather Taihang cock,Taihang hen and fast-feather Taihang cock was 94.1%,65.5% and0.0%,respectively,and it was 100.0% and 0.0%in slow-feather and fast-feather progenitors of Hyline gray,respectively.The 1 440 bp fragment could not be digested by HaeⅢ and it could be a detection basis for ev21.The structure was analyzed comprehensively for the OS and US of ev21 in 6strains of chicken and it was concluded that the site recognized by Hae Ⅲ enzyme in US could not be a detection base for slow-feather and ev21,while ev21 insertion in OS and the site for HaeⅢ digestion in 1 440 bp were linked with the mutation of A → G and 8bases repeated.更多还原