本文選題：鵝 + GnRH�。� 參考：《西南大學(xué)》2017年碩士論文

【摘要】：產(chǎn)蛋性狀是鵝重要的經(jīng)濟(jì)性狀,而鵝產(chǎn)蛋性能低下嚴(yán)重制約鵝產(chǎn)業(yè)的發(fā)展。禽類的生殖功能是由下丘腦-垂體-性腺軸系統(tǒng)的各類激素之間復(fù)雜的相互調(diào)控作用所決定。GnRH與GnIH已被證明是下丘腦-垂體-性腺軸(hypothalamus-pituitary-gonadal axis,HPG)的關(guān)鍵信號分子。下丘腦釋放的促性腺激素釋放激素與抑制激素作用于垂體,調(diào)節(jié)垂體促性腺激素的分泌,促性腺激素通過外周血液循環(huán)影響性腺的內(nèi)分泌和外分泌,從而控制動物的個體發(fā)育以及生殖過程。有研究表明,GnRH與GnIH基因可作為影響豬、牛、羊等動物繁殖性能的候選基因。目前,有關(guān)鵝GnRH與GnIH基因的研究很少見報道。本研究選用四川白鵝為試驗動物,開展了鵝GnRH和GnIH基因組序列克隆、基因結(jié)構(gòu)分析與SNP檢測;采用PCR-RFLP、PCR-SSCP的方法檢測鵝GnRH、GnIH的基因型并分析其與產(chǎn)蛋量的關(guān)聯(lián)性。獲得的主要研究結(jié)果如下:四川白鵝GnRH基因DNA序列長3520bp,包括4個外顯子和3個內(nèi)含子;mRNA序列比對發(fā)現(xiàn)鵝與雁GnRH基因的相似性最高,達(dá)到92%;共檢測到鵝GnRH基因28個SNPs位點,包括外顯子上的1個SNP位點(g.3447AC)和內(nèi)含子上的27個SNPs位點;對內(nèi)含子1的突變位點(g.490AG)采用BclⅠ限制性內(nèi)切酶進(jìn)行酶切分型,共產(chǎn)生3種基因型,分別定義為AA、AB、BB,三種基因型鵝的產(chǎn)蛋量無顯著差異(P0.05);對外顯子的突變位點(g.3447TG)用PCR-SSCP進(jìn)行分析,得到三種基因型(TT、TG、GG),三種基因型鵝的產(chǎn)蛋量差異不顯著(P0.05)。四川白鵝GnIH基因DNA序列長7621bp,包括4個外顯子和3個內(nèi)含子;mRNA序列比對發(fā)現(xiàn)鵝GnIH基因與灰雁、原雞、鵪鶉的相似性分別高達(dá)99%、91%、90%。檢測到鵝GnIH基因內(nèi)含子上有18個SNPs,外顯子上有15個SNPs,其中外顯子230bp處的堿基GAA缺失、2164bp處的CG突變、2228bp處的TA突變均導(dǎo)致氨基酸發(fā)生改變,分別為p.38Glu In Del、p.100ProArg、p.123TryAsn。采用PCR-SSCP方法對外顯子1的230bp處堿基GAA缺失與內(nèi)含子2的3262bp處堿基TTTTG缺失進(jìn)行基因型分析,均產(chǎn)生兩種基因型,分別定義為AA、Aa型與HH、Hh型;AA、Aa基因型及HH、Hh基因型鵝產(chǎn)蛋量差異均不顯著(P0.05)。采用SmaⅠ與EcoR V酶切的方法分別對內(nèi)含子1(g.1320GA)與外顯子3(g.3553TC)的突變位點進(jìn)行基因型分析,結(jié)果顯示:1320bp處酶切產(chǎn)生的CC基因型鵝產(chǎn)蛋量顯著高于CD與DD基因型(P0.05);3553bp酶切產(chǎn)生的EE、EF、FF三種基因型鵝產(chǎn)蛋量差異不顯著(P0.05)。本研究首次獲得了四川白鵝GnRH、GnIH基因DNA序列與SNP位點;GnIH基因g.1320GA突變所產(chǎn)生的CC基因型產(chǎn)蛋量顯著高于其他兩種基因型產(chǎn)蛋量。該研究結(jié)果為鵝GnRH、GnIH基因功能的進(jìn)一步研究奠定基礎(chǔ),為鵝產(chǎn)蛋調(diào)控機制的研究提供了參考。
[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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