慢羽系麒麟公鸡纯合个体分子检测方法的建立  

Establishment of Molecular Identification Techniques for Frizzle Chicken Early Feathering and Late Feathering

作　　者：李珊珊[1] 李东华[1] 吕福琨 董晶[1] 李艳青[1] 杜炳旺[1] 

机构地区：[1]广东海洋大学动物科学系,广东湛江524088

出　　处：《河南农业科学》2016年第7期118-121,共4页Journal of Henan Agricultural Sciences

基　　金：广东省创新强校重大项目(GDOU2013050222)

摘　　要：为建立一种快速准确鉴定麒麟公鸡慢羽纯合子的分子检测方法,以加快纯合慢羽系及其配套系的育种速度,利用鸡性连锁羽速基因K/k中慢羽基因K与禽白血病内源性病毒基因ev21的紧密连锁关系,选择241只(直观鉴定171只为表型慢羽,70只为表型快羽)140日龄麒麟公鸡,应用PCR扩增URa和URb基因以鉴定其快、慢羽型,再扩增ev21基因,并采用限制性内切酶HaeⅢ对慢羽鸡扩增产物进行消化,检测基因型为纯合子的慢羽公鸡。麒麟鸡羽速类型检测结果显示,慢羽公鸡(170只)2条带,快羽麒麟公鸡(71只)1条带。慢羽公鸡酶切基因分型结果显示,酶切后有2条带的为ev21缺失个体(72只),有3条带的是杂合个体(94只),有1条带的为纯合个体(5只)。综上,可以利用ev21插入位点鉴定麒麟鸡基因型是否为纯合的慢羽个体。In order to establish a rapid and accurate molecular identification techniques for the frizzle chicken early feathering and late feathering to speed up the formation of complete set line,male Kirin chickens of 241 at 140 days were selected（ 171 late-feathering phenotypic,70 eaily-feathering phenotypic）.The slight distance relation of late-feathering gene K in linkage with feathering gene K / k of chicken and endogenous ev21 was used, to discriminate the eaily-feathering and late-feathering cock using PCR amplification of ev21 gene and to detect homozygote of genotype late-feathering cock through the amplified products digestion using the restriction enzyme（ HaeⅢ）. The results showed that eaily-feathering frizzles had a band（ 71）,late-feathering had two bands（ 170）. Compared with eye identification,the agreement rate was99. 41%. The Hae Ⅲ cleaved genotyping results of late-feathering cock showed that the chicken missing ev21 had two bands（ 72）,the individuals with three bands were heterozygous（ 94）,and homozygous individuals had a band（ 5）. Therefore,ev21 insertion site could be used for identifying genotype homozygous Kirin male chicken.

关 键 词：麒麟鸡 羽速基因 ev21基因 纯合子 配套系 

分 类 号：S831[农业科学—畜牧学]