赤水乌骨鸡MC1R基因多态性及其生物信息学分析

Polymorphism and bioinformatics analysis of MC1R gene in Chishui Black-Bone chickens

采用DNA混池及PCR产物直接测序技术,对赤水乌骨鸡、泰和乌鸡、兴义白鸡、兴义白鸡F2代、贵州黄快羽鸡、贵州黄慢羽鸡、罗曼蛋鸡和瑶山鸡8个鸡种群MC1R基因外显子区域进行多态性分析。结果显示:瑶山鸡与红色原鸡序列一致,无SNPs,赤水乌骨鸡的4个SNPs分别为G23A(Arg→His)、C69T、T212C、G274A,泰和乌鸡的8个SNPs分别为C69T、T212C、G274A、T398C(Leu→Pro)、G636A、T637C、A644C(His→Pro)、C834T,贵州黄快、慢羽鸡和兴义白鸡均有3个SNPs,分别为A427G(Thr→Ala)、G636A、T637C,罗曼蛋鸡的4个SNPs分别为T398A(Leu→Gln)、G636A、T637C、C834T,兴义白鸡F2代的4个SNPs分别为C69T、T212C、G274A、A644C(His→Thr);生物信息学分析发现,除G274A、T398C、T398A、A644C位点的稳定性增加外,其余位点的稳定性均有所降低;除G274A、T398C、T637C、A644C位点的整体多样性有所下降外,其余位点的均有所增加;除泰和乌鸡MC1R基因编码蛋白为不稳定蛋白外,其余种群的均属于稳定蛋白,且包括泰和乌鸡在内的7个鸡群MC1R基因编码蛋白均为疏水性蛋白和非分泌蛋白;赤水乌骨鸡、泰和乌鸡、兴义白鸡、兴义白鸡F2代和罗曼蛋鸡的MC1R蛋白三级结构均由α-螺旋、β-转角、无规则卷曲组成。

DNA mixing pool and direct sequencing of PCR products were used to analyze the polymorphism of MC1 R exon regions in 8 chicken populations of Chishui Black-Bone Chickens,Taihe Silky Fowl,Xingyi White Chickens,the F2 Xingyi White Chickens,Guizhou Yellow Fast Chickens,Guizhou Yellow Slow Chickens,Luoman Laying Hens and Yaoshan Chickens.The results showed that the sequence of Yaoshan Chickens was identical to that of Red Jungle Fowl without SNPs.The four SNPs of Chishui Black-Bone Chickens were G23 A(Arg→His),C69 T,T212 C and G274 A.The eight SNPs of Taihe Silky Fowl were C69 T,T212 C,G274 A,T398 C(Leu→Pro),G636 A,T637 C,A644 C(His→Pro)and C834 T.Guizhou Yellow Fast Chickens,Guizhou Yellow Slow Chickens and Xingyi White Chickens all had 3 SNPs,A427 G(Thr→Ala),G636 A and T637 C.The 4 SNPs of Luoman Laying Hens were T398 A(Leu→Gln),G636 A,T637 C and C834 T.The 4 SNPs of the F2 Xingyi White Chickens were C69 T,T212 C,G274 A and A644 C(His→Thr).The bioinformatics analysis found that except for the stability of G274 A,T398 C,T398 A and A644 C sites,the stability of the other sites decreased.Except that Taihe Silky Fowl,whose MC1 R gene encoding protein was unstable,the other populations’s MC1 R protein were stable,and all the proteins from the 7 chicken populations were owing hydrophobic and non-secreted character.The tertiary structure of MC1 R protein of Chishui Black-Bone Chickens,Taihe Silky Fowl,Xingyi White Chickens,the F2 Xingyi White Chickens and Luoman Laying Hens were composed ofα-helix,β-turn,and random curl.