文昌鸡产蛋性能及蛋品质性状分子标记的研究
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摘要
以文昌鸡作为试验样本,采用候选基因法,以GNRHR、NPY、GHR (intron 2、intron 5)、PRL、ESR462、ESR465、IGF-I、TGFβ-2、TGFβ-3、TGFβ-4十一个基因座位候选基因,以及鸡2号、4号、Z染色体上20个微卫星标记进行鸡产蛋性能和蛋品质性状分子标记研究。
1.首次在文昌鸡中检测了与产蛋性能相关的十一个候选基因座位的多态性。在这十一个基因位点中,处于Hardy-Wenberg平衡状态的有六个,它们分别是GNRHR (intron 1)、NPY (TSS)、PRL450 (intron 1)、ESR462 (5' flanking region)、IGF (5′UTR)、TGFβ-3 (intron 4);非Hardy-Wenberg平衡状态的有五个:GHR442 (intron 2)、GHR436 (intron 5)、ESR465 (5' flanking region)、TGFβ-2 (Promoter region)、TGFβ-4(exon 1)。其中ESR465 (5' flanking region)、TGFβ-2 (Promoter region)、TGFβ-4(exon 1)位点中未发现多态。
2.初步确定了1个新的与鸡产蛋性能相关的分子标记,该标记位于ESR基因5'调控区域,对两种纯合基因型的片段进行回收、克隆、测序比较发现,ESR基因5’侧翼序列片段中第580位核苷酸AATA缺失,因而造成该基因5’侧翼序列的多态性。
确定了4个与鸡产蛋性能相关的分子标记:GNRHR基因内含子1处的Bpu1102I酶切位点、NPY基因转录起始区域的DraI酶切位点、ESR基因5'调控区域的SSCP位点、IGF-I基因5'非编码区域PstI酶切位点;3个与鸡蛋品质性状相关的分子标记:GHR基因内含子5的NspI酶切位点; IGF-I基因5'非编码区域PstI酶切位点;TGFβ-3内含子4的BslI酶切位点。
Egg production and egg quality traits of 120 hens from a pureline of Wenchang layers (Chinese indigenous chicken breed) were collected in testing center of poultry quality, Ministry Agriculture of China. Data on egg production including total egg production, continual egg-laying, and number of double-yolked eggs were collected daily using trap nests to identify individual bird. The data for individual hen were collated over a 5-month period and recording commenced at 25 weeks of age and all birds were treated the same. Egg quality including egg weight(EW)、shell weight (SW)、shell thickness (ST)、shell strength (SS)、haugh unit (HU)、egg shape index (SI)、yolk weight (YW) and yolk color (YC) were measured at the age of 55 weeks.
A candidate gene approach was used to determine whether specific loci explain responses in the egg production and egg quality traits of Wenchang chicken in its pure-line population. PCR-RFLP and PCR-SSCP techniques were conducted on nine candidate genes that were gonadotropin releasing hormone receptor (GNRHR)、Neuropeptide Y(NPY)、Growth hormone receptor(GHR)、prolactin(PRL)、Estrogen Receptor (ESR)、insulin-like growth factor-1(IGF-I)、transforming growth factorβ-2(TGFβ-2)、transforming growth factorβ-3(TGFβ-3)and transforming growth factorβ-4(TGFβ-4)respectively. And microsatellite primers of 20 pairs in total have been employed to perform electrophoresis to determine the genotype of individual. This study was designed to identify the polymorphisms of each gene and microsatellite locus and found genetic effects on the traits. The results is as following.
Polymorphic sites were found in each genes except for TGFβ-2 and TGFβ-4 andχ~2 analysis among these gene locus indicated that only the frequencies of GNRHR、NPY、PRL、ESR、IGF-I and TGFβ-3 fit Hardy-Weinberg equilibrium. It suggested that allele and genotypes frequencies of these sites were stable. In addition,association analysis of the
1.首次在文昌鸡中检测了与产蛋性能相关的十一个候选基因座位的多态性。在这十一个基因位点中,处于Hardy-Wenberg平衡状态的有六个,它们分别是GNRHR (intron 1)、NPY (TSS)、PRL450 (intron 1)、ESR462 (5' flanking region)、IGF (5′UTR)、TGFβ-3 (intron 4);非Hardy-Wenberg平衡状态的有五个:GHR442 (intron 2)、GHR436 (intron 5)、ESR465 (5' flanking region)、TGFβ-2 (Promoter region)、TGFβ-4(exon 1)。其中ESR465 (5' flanking region)、TGFβ-2 (Promoter region)、TGFβ-4(exon 1)位点中未发现多态。
2.初步确定了1个新的与鸡产蛋性能相关的分子标记,该标记位于ESR基因5'调控区域,对两种纯合基因型的片段进行回收、克隆、测序比较发现,ESR基因5’侧翼序列片段中第580位核苷酸AATA缺失,因而造成该基因5’侧翼序列的多态性。
确定了4个与鸡产蛋性能相关的分子标记:GNRHR基因内含子1处的Bpu1102I酶切位点、NPY基因转录起始区域的DraI酶切位点、ESR基因5'调控区域的SSCP位点、IGF-I基因5'非编码区域PstI酶切位点;3个与鸡蛋品质性状相关的分子标记:GHR基因内含子5的NspI酶切位点; IGF-I基因5'非编码区域PstI酶切位点;TGFβ-3内含子4的BslI酶切位点。
Egg production and egg quality traits of 120 hens from a pureline of Wenchang layers (Chinese indigenous chicken breed) were collected in testing center of poultry quality, Ministry Agriculture of China. Data on egg production including total egg production, continual egg-laying, and number of double-yolked eggs were collected daily using trap nests to identify individual bird. The data for individual hen were collated over a 5-month period and recording commenced at 25 weeks of age and all birds were treated the same. Egg quality including egg weight(EW)、shell weight (SW)、shell thickness (ST)、shell strength (SS)、haugh unit (HU)、egg shape index (SI)、yolk weight (YW) and yolk color (YC) were measured at the age of 55 weeks.
A candidate gene approach was used to determine whether specific loci explain responses in the egg production and egg quality traits of Wenchang chicken in its pure-line population. PCR-RFLP and PCR-SSCP techniques were conducted on nine candidate genes that were gonadotropin releasing hormone receptor (GNRHR)、Neuropeptide Y(NPY)、Growth hormone receptor(GHR)、prolactin(PRL)、Estrogen Receptor (ESR)、insulin-like growth factor-1(IGF-I)、transforming growth factorβ-2(TGFβ-2)、transforming growth factorβ-3(TGFβ-3)and transforming growth factorβ-4(TGFβ-4)respectively. And microsatellite primers of 20 pairs in total have been employed to perform electrophoresis to determine the genotype of individual. This study was designed to identify the polymorphisms of each gene and microsatellite locus and found genetic effects on the traits. The results is as following.
Polymorphic sites were found in each genes except for TGFβ-2 and TGFβ-4 andχ~2 analysis among these gene locus indicated that only the frequencies of GNRHR、NPY、PRL、ESR、IGF-I and TGFβ-3 fit Hardy-Weinberg equilibrium. It suggested that allele and genotypes frequencies of these sites were stable. In addition,association analysis of the
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