ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2016, Vol. 47 ›› Issue (12): 2354-2361.doi: 10.11843/j.issn.0366-6964.2016.12.004

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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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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.

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