四川白鹅GnRH、GnIH基因克

发布时间：2018-06-30 04:07

  本文选题：鹅 + GnRH　； 参考：《西南大学》2017年硕士论文

【摘要】：产蛋性状是鹅重要的经济性状,而鹅产蛋性能低下严重制约鹅产业的发展。禽类的生殖功能是由下丘脑-垂体-性腺轴系统的各类激素之间复杂的相互调控作用所决定。GnRH与GnIH已被证明是下丘脑-垂体-性腺轴(hypothalamus-pituitary-gonadal axis,HPG)的关键信号分子。下丘脑释放的促性腺激素释放激素与抑制激素作用于垂体,调节垂体促性腺激素的分泌,促性腺激素通过外周血液循环影响性腺的内分泌和外分泌,从而控制动物的个体发育以及生殖过程。有研究表明,GnRH与GnIH基因可作为影响猪、牛、羊等动物繁殖性能的候选基因。目前,有关鹅GnRH与GnIH基因的研究很少见报道。本研究选用四川白鹅为试验动物,开展了鹅GnRH和GnIH基因组序列克隆、基因结构分析与SNP检测;采用PCR-RFLP、PCR-SSCP的方法检测鹅GnRH、GnIH的基因型并分析其与产蛋量的关联性。获得的主要研究结果如下:四川白鹅GnRH基因DNA序列长3520bp,包括4个外显子和3个内含子;mRNA序列比对发现鹅与雁GnRH基因的相似性最高,达到92%;共检测到鹅GnRH基因28个SNPs位点,包括外显子上的1个SNP位点(g.3447AC)和内含子上的27个SNPs位点;对内含子1的突变位点(g.490AG)采用BclⅠ限制性内切酶进行酶切分型,共产生3种基因型,分别定义为AA、AB、BB,三种基因型鹅的产蛋量无显著差异(P0.05);对外显子的突变位点(g.3447TG)用PCR-SSCP进行分析,得到三种基因型(TT、TG、GG),三种基因型鹅的产蛋量差异不显著(P0.05)。四川白鹅GnIH基因DNA序列长7621bp,包括4个外显子和3个内含子;mRNA序列比对发现鹅GnIH基因与灰雁、原鸡、鹌鹑的相似性分别高达99%、91%、90%。检测到鹅GnIH基因内含子上有18个SNPs,外显子上有15个SNPs,其中外显子230bp处的碱基GAA缺失、2164bp处的CG突变、2228bp处的TA突变均导致氨基酸发生改变,分别为p.38Glu In Del、p.100ProArg、p.123TryAsn。采用PCR-SSCP方法对外显子1的230bp处碱基GAA缺失与内含子2的3262bp处碱基TTTTG缺失进行基因型分析,均产生两种基因型,分别定义为AA、Aa型与HH、Hh型;AA、Aa基因型及HH、Hh基因型鹅产蛋量差异均不显著(P0.05)。采用SmaⅠ与EcoR V酶切的方法分别对内含子1(g.1320GA)与外显子3(g.3553TC)的突变位点进行基因型分析,结果显示:1320bp处酶切产生的CC基因型鹅产蛋量显著高于CD与DD基因型(P0.05);3553bp酶切产生的EE、EF、FF三种基因型鹅产蛋量差异不显著(P0.05)。本研究首次获得了四川白鹅GnRH、GnIH基因DNA序列与SNP位点;GnIH基因g.1320GA突变所产生的CC基因型产蛋量显著高于其他两种基因型产蛋量。该研究结果为鹅GnRH、GnIH基因功能的进一步研究奠定基础,为鹅产蛋调控机制的研究提供了参考。
[Abstract]:Egg-laying character is an important economic trait of geese, and the low egg-laying performance of geese seriously restricts the development of goose industry. The reproductive function of poultry is determined by the complex regulation of hormones in the hypothalamus-pituitary-gonadal axis system. GnRH and GnIH have been proved to be the key signal molecules of the hypothalamus-pituitary-gonadal axis-gonadal axis (HPG). Hypothalamic gonadotropin releasing hormone and inhibiting hormone act on the pituitary gland and regulate the secretion of pituitary gonadotropin. Gonadotropin affects the endocrine and exocrine secretion of gonad through peripheral blood circulation. Thus controlling the ontogeny and reproduction of animals. Some studies have shown that GnRH and GnIH genes can be used as candidate genes to affect the reproductive performance of pigs, cattle, sheep and other animals. At present, the studies of goose GnRH and GnIH genes are rarely reported. In this study, Sichuan White Goose was selected as experimental animal to carry out genomic sequence cloning of goose GnRH and GnIH, gene structure analysis and SNP detection, PCR-RFLP- PCR-SSCP method to detect the genotype of goose GnRHH GnIH and to analyze its correlation with egg production. The main results were as follows: the length of GnRH gene DNA sequence of Sichuan White Goose was 3520bp. the sequence alignment of GnRH gene including 4 exons and 3 introns showed that the similarity of GnRH gene between goose and Goose was the highest, which reached 92k.A total of 28 SNPs loci of goose GnRH gene were detected. There were one SNP locus on exon (g.3447AC) and 27 SNPs loci on intron, and three genotypes were produced by Bcl 鈪，

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