寿光鸡AANAT基因5’调控区的多态性及其与产蛋性状的关联分析

发布时间：2018-04-25 22:37

本文选题：寿光鸡 + 褪黑激素AANAT基因　；参考：《山东农业大学》2015年硕士论文

【摘要】：褪黑激素主要是由哺乳动物和人类的松果体产生的一种吲哚类化合物,具有广泛的生理活性。褪黑激素的合成分四步进行,以色氨酸为原料,先后经过羟化,脱羧,N2乙酰化和氧甲基化,最终形成褪黑激素。无论两栖类、爬行类、哺乳动物还是鸟类,其合成步骤是这样的。合成过程中涉及两个主要的酶,AANAT酶(5-羟色胺-N-乙酰基转移酶)是限速酶。HIOMT是关键酶。本研究拟采用实验性的方法对AANAT基因5'调控区多态性和鸡产蛋性状进行关联分析,旨在发现有意义的分子标记,辅助实际蛋鸡育种工作,同时期望AANAT基因的研究对完善褪黑激素作用的分子机制能有一定贡献。本实验以寿光鸡为材料,对鸡的AANAT基因5'调控区进行PCR扩增,再用单克隆测序得到AANAT基因5'调控区可能存在的多态性位点,然后用直接测序的方法大群体检测其多态性位点,并将各多态性位点与记录的寿光鸡产蛋性状进行关联分析,分析的部分结果如下:AANAT基因5'调控区的共有24个SNPs位点。其中,15处碱基转换、7处碱基颠换和2处碱基插入。-853、-803、-623、-619、-561、-522、-477、-210共8个位点对鸡的产蛋性能有显著影响(P0.05),其他突变SNPs位点对产蛋性能均无显著影响。其中,4个低度多态位点-853、-623、-619、-477位点,其不同基因型的频率差距太大,个别基因型频率特别小,代表性差。-803位点等位基因G比T能使鸡提前开产约2.5天,早期产蛋数增加约1.6枚,该位点对开产日龄和早期产蛋数的作用方式为加性效应和显性效应;-561位点等位基因T比A能使鸡中期产蛋数增加0.9枚,等位基因T比A能使鸡产蛋总数增加4.4枚,对中期产蛋数的作用方式主要为显性效应,加性效应不显著。对于产蛋总数,等位基因T比A能使鸡产蛋总数增加4.4枚,作用方式为加性效应和显性效应,均显著;-522位点等位基因A比G能使鸡后期产蛋数增加1.5枚,等位基因A比G能使鸡产蛋总数增加5.4枚,该位点对后期产蛋数和产蛋总数的作用方式为加性效应和显性效应,且均显著;-210位点等位基因A比C能使鸡提前2.9天开产,产蛋总数增加5.8枚。-708和-615位点连锁,构建了4种单倍型,与单倍型TG相比,TA单倍型使母鸡的开产日龄延后了10.3天(P0.05);-619和-595位点连锁,构建了4种单倍型,与单倍型GT相比,单倍型GC能使母鸡的中期产蛋数提高10.6枚(P0.05),单倍型AC和GC之间还存在显著的互作效应,使早期产蛋数和中期产蛋数减少;-546和-210位点连锁,构建了4种单倍型,与单倍型TA相比,单倍型CC使母鸡的开产日龄提前了约4.0天(P0.05),而单倍型CA使母鸡的后期产蛋数减少了约6.8枚(P0.05)。本研究发现一些潜在的分子标记,为寿光鸡的遗传选育改良及褪黑激素受体调控鸡繁殖性能的分子机制提供参考。
[Abstract]:Melatonin is an indole compound produced mainly by the pineal gland of mammals and humans and has a wide range of physiological activities. The synthesis of melatonin was divided into four steps. Using tryptophan as raw material, melatonin was formed by hydroxylation, decarboxylation of N _ 2 acetylation and oxygen methylation. Whether amphibians, reptiles, mammals or birds, the synthesis process is like this. The rate-limiting enzyme, HIOMT, is the key enzyme involved in the synthesis of two major enzymes, AANAT and 5-hydroxytryptamine-N-acetyltransferase. The aim of this study was to use experimental methods to analyze the polymorphism of 5'regulatory region of AANAT gene and egg laying traits in order to find meaningful molecular markers to assist the breeding of laying hens. At the same time, it is expected that the study of AANAT gene will contribute to the improvement of the molecular mechanism of melatonin. The 5 'regulatory region of AANAT gene was amplified by PCR from Shouguang chicken. The polymorphic loci of 5' regulatory region of AANAT gene were obtained by monoclonal sequencing, and the polymorphic loci of 5 'regulatory region of AANAT gene were detected by direct sequencing method. The polymorphic loci were associated with egg laying traits recorded in Shouguang Chicken. Some of the results were as follows: a total of 24 SNPs loci were located in the 5 'regulatory region of the 1: AANat gene. Among them, 15 base transversion sites and 2 base inserts. -853An -803U -623C -619U -561U -522U -477C -210 had significant effects on laying performance of chickens, while other mutant SNPs sites had no significant effect on egg laying performance. Among them, four low-degree polymorphic loci -853r-623C-619C-477, the frequency difference of different genotypes was too big, and the frequency of individual genotypes was very small. The allele G of .-803 locus could make chickens produce about 2.5 days earlier than T, and increase the number of eggs in the early stage by about 1.6. The effect of this locus on laying age and early egg number was as follows: additive effect and dominant effect: allele T and dominant effect could increase the number of eggs produced in the middle period of chicken by 0.9 alleles than A, and allele T ratio A could increase the total number of eggs produced by chicken by 4.4. The dominant effect was dominant effect, but the additive effect was not significant. For the total number of eggs laid, allele T than A could increase the total number of eggs by 4.4, in the form of additive effect and dominant effect, and allele A at locus 522 significantly increased the number of eggs laid at the later stage by 1.5 compared with G. Allele A could increase the total number of eggs by 5.4 than G. The effect of allele A on the number of eggs and the total number of eggs in the later stage was additive and dominant, and allele A at locus A was significantly higher than that at locus C by 2.9 days earlier than that of C. The total number of eggs was increased by 5.8 loci of .-708 and -615, and four haplotypes were constructed. Compared with the haplotype TG, TA haplotype delayed the age of birth of hens by 10.3 days (P 0.05) -619 and -595, and constructed four haplotypes, which were compared with haplotype GT. Haplotype GC could increase the egg number of hens by 10.6 eggs in the middle stage. There was also a significant interaction effect between haplotype AC and GC, which reduced the number of eggs laid in the early stage and the number of eggs laid in the metaphase by loci -546 and -210. Four haplotypes were constructed, which were compared with haplotype TA. Haplotype CC could advance the age of laying hen by 4.0 days, and haplotype CA could reduce the number of eggs laid by hens at later stage. In this study, some potential molecular markers were found to provide reference for the genetic improvement of Shouguang chicken and the molecular mechanism of melatonin receptor regulating chicken reproductive performance.
【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S831

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