畜牧兽医学报 ›› 2016, Vol. 47 ›› Issue (12): 2354-2361.doi: 10.11843/j.issn.0366-6964.2016.12.004

• 遗传繁育 • 上一篇    下一篇

鸡GNAS基因启动子突变及其与肤色性状的相关性

王欢欢1，陈美玲1，楼立峰1，张雷1，张成先2，陈贤惠2，章学东1*   

  1. （1. 杭州市农业科学研究院，杭州 310024； 2. 浙江光大种禽业有限公司，海宁 310016）
  • 收稿日期:2016-05-16 出版日期:2016-12-23 发布日期:2016-12-23
  • 通讯作者: 章学东，研究员，E-mail：bigzhengliang@hotmail.com
  • 作者简介:王欢欢(1986-)，女，黑龙江佳木斯人，畜牧师，硕士，主要从事动物遗传育种工作，E-mail: wennuan2009@163.com
  • 基金资助:

    国家肉鸡产业技术体系综合试验站（CARS-42-Z07）；浙江省公益技术研究农业项目（2015C32106）

GNAS Gene Promoter Mutation in Chicken and the Correlation with Skin Color Traits

WANG Huan-huan1，CHEN Mei-ling1，LOU Li-feng1，ZHANG Lei1，ZHANG Cheng-xian2，CHEN Xian-hui2，ZHANG Xue-dong1*   

  1. （1. Hangzhou Academy of Agricultural Sciences，Hangzhou 310024，China；  2.Zhejiang Guangda Poultry Breeding Company，Haining 310016，China）
  • Received:2016-05-16 Online:2016-12-23 Published:2016-12-23

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摘要：

为研究鸡GNAS基因突变情况及其对乌骨鸡肤色的影响，本试验选择东乡绿壳蛋鸡样本，采用PCR-RFLP和DNA测序方法，筛查肤色相关的GNAS突变位点；采用荧光定量PCR方法，检测EDN3、MC1R、GNAS、ADCY3基因的表达量；本试验还对3个鸡种进行了突变检测。结果显示，在家鸡20号染色体的11 167 660碱基位置存在T/C突变，该T2270C突变点是GNAS基因启动子的关键位点。PCR产物经Dra I酶切电泳后，CC纯合基因型出现一条342 bp的条带；TC杂合基因型出现171和342 bp两条带；TT隐性纯合基因型出现一条171 bp的条带。3种基因型的肤色L值差异极显著（P＜0.01），CC基因型的L值最小，表现为肤色最深；TT基因型的L值最大，表现为肤色浅白。EDN3基因的表达量以CC基因型最高，TT基因型最低，3种基因型间差异极显著（P＜0.01）。MC1R、GNAS和ADCY3基因的表达量以CC基因型最高，但基因型间的表达差异均不显著（P＞0.05）；GNAS与上游MC1R基因的表达量无显著相关性，而与下游ADCY3基因的表达量极显著相关（P＜0.01）。江山乌骨鸡、伊莎蛋鸡、东乡×伊莎测交组合鸡的突变分型结果符合理论值（P＞0.05）。本研究表明，鸡GNAS基因启动子T2270C突变与肤色性状强相关，突变检测可辅助用于乌骨鸡的肤色分型育种。

Abstract:

The study aimed to investigate the mutation of GNAS gene and its effect on skin color traits in chicken. Dongxiang Blue-eggshell chicken (Gallus gallus domestica) was chose and the GNAS mutation sites related to skin color were screened using PCR-restriction length polymorphism (RFLP) and DNA sequencing in this study. We detected expression of the EDN3, MC1R, GNAS and ADCY3 genes using fluorescence quantitative PCR, and the mutation sites among 3 chicken breeds. The results showed that there was a T/C mutation at base 11 167 660 on chicken chromosome 20 and the mutation (T2270C) was crucial for the GNAS gene promoter. After Dra I enzyme digestion for PCR products, the CC homozygous genotype, the TC heterozygous genotype and the TT recessive homozygous genotype presented one band at 342 bp, two bands at 171 and 342 bp, and one band at 171 bp, respectively. The L values of skin color were statistically significantly different among 3 genotypes (P＜0.01). The individuals with CC genotype had the lowest L value, which presented the darkest skin, and the individuals with TT genotype had the highest L value, which presented the lightest skin. Expression of the EDN3 gene was highest and lowest in individuals with the CC and TT genotype, respectively, and the difference among 3 genotypes was extremely significant (P＜0.01). Expression of the MC1R, GNAS and ADCY3 genes was highest in individuals with CC genotype, but the difference among genotypes was not significant (P＞0.05). The correlation of GNAS with MC1R was non-significant in expression, and the correlation with ADCY3 was extremely significant (P＜0.01). The typing results according to mutation detection in Jiangshan Black-bone chicken, ISA layer, test-crossing offspring of Dongxiang Blue-eggshell cock and ISA hen, conformed to the theoretical values (P＞0.05). The study indicate that the T2270C mutation in GNAS gene promoter in chicken is correlated strongly with the skin color traits, and detection of the mutation can be used in the breeding of black-bone chicken.

中图分类号: 

ISSN 0366-6964
CN 11-1985/S
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