京海黄鸡FSHβ和FSHR基因的遗传效应及其表达规律研究

发布时间：2018-05-29 10:01

本文选题：京海黄鸡 + FSHβ基因　；参考：《扬州大学》2015年硕士论文

【摘要】：传统育种方法在家禽的很多重要经济性状中取得了巨大的进展,但耗时耗力。近些年来,在家禽育种中采用了分子生物技术,候选基因法是一种较常用的方法,以期找到与经济性状变异相关的数量性状座位(quantitative trait locus, QTL)。作为重要的经济性状,体重和繁殖性状都是由繁杂的遗传和环境因素控制,所以研究家禽生长和繁殖的分子机制,可用于选择家禽重要的经济性状。本研究采用PCR-SSCP技术检测京海黄鸡促卵泡激素(follicle-stimulating hormone, FSH)β亚基基因及其受体(follicle-stimulating hormone receptor, FSHR)基因的多态性,并研究SNPs与生产性状的关系；使用RQ-PCR法研究京海黄鸡FSHβ和FSHR基因的表达谱及表达规律。主要研究结果如下：1.检测FSHβ基因的全部外显子,分析其单核苷酸多态性。结果显示FSHβ基因仅外显子3存在多态性,其中相邻的C2049T和A2050T突变,形成AA、AB和BB 3种基因型；T2341C突变形成CC、CD和DD 3种基因型；A2467C突变形成EE、EF和FF 3种基因型。关联分析结果显示12周龄体重AB基因型显著高于BB基因型(P0.05)、EF基因型显著高于EE基因型(P0.05)；2周龄体重CC基因型极显著高于CD基因型(P0.01)；并且以上突变对繁殖性状无显著影响。因而可知,在生长性状方面,AB、CC和EF基因型为京海黄鸡的优势基因型,可初步推断FSHβ基因对京海黄鸡的生长有一定的促进作用。2.检测FSHR基因的外显子1和10,分析其单核苷酸多态性。结果显示FSHR基因外显子1处存在C159T突变,形成AA、AB和BB三种基因型；FSHR基因外显子10处存在C75429T的点突变,形成三种基因型CC、CD和DD。关联分析结果显示京海黄鸡2周龄体重AB基因型极显著高于BB基因型(P0.01)、AA基因型显著高于BB基因型(P0.05)；6周龄体重AA和AB基因型都显著高于BB基因型(P0.05)、CD基因型显著高于CC基因型(P0.05)；开产日龄CC和CD基因型显著早于DD基因型(P0.05)；300日龄产蛋数AB基因型显著高于AA基因型(P0.05)；其余差异不显著。3.对京海黄鸡FSHβ基因在不同生长阶段和组织中的表达规律进行研究。结果显示FSHβ基因在不同组织中均有一定的表达量,其中性腺和垂体的表达水平最高,在0-20周龄,FSHβ基因nRNA表达丰度在20周。FSHβ基因在公母鸡上的表达趋势是一样的,在垂体中初生时相对表达量几乎为零,到4周龄有小幅度上升的趋势,从4周龄到16周龄表达量有很大的提高,母鸡20周龄达到最高,即从4周龄到20周龄FSHβ基因的表达呈直线上升。在卵巢中初生时有少量表达,然后到4周龄降到最低之后趋于稳定。另外,4周龄和16周龄时FSHβ基因在母鸡性腺和垂体中的相对表达量均高于公鸡。4.对京海黄鸡FSHR基因在不同生长阶段和组织中的表达规律进行研究。结果显示FSHR基因在性腺表达量最高,表达丰度在初生时。在0-20周龄,FSHR基因在卵巢和睾丸中的表达,在4周龄、16周龄时都有分别下降的趋势,20周龄母鸡卵巢中FSHR基因表达量较16周龄有略微的上升,随着日龄的增长而降低。在垂体上,0-20周龄中,FSHR基因在京海黄鸡上的表达缓慢平稳的有小幅度的上升,相对来说,同一时期的公母鸡垂体中FSHR基因的表达量相仿,母鸡卵巢的表达量稍高于公鸡睾丸中的表达量。
[Abstract]:The traditional breeding method has made great progress in many important economic traits of poultry, but time consuming. In recent years, molecular biotechnology is used in poultry breeding. Candidate gene method is a more common method to find quantitative trait locus (QTL), which is related to the variation of economic traits. Important economic traits, weight and reproductive traits are controlled by complex genetic and environmental factors, so the molecular mechanism of poultry growth and reproduction can be used to select important economic traits in poultry. This study uses PCR-SSCP technology to detect the follicle-stimulating hormone (FSH) beta subunit gene and its subunit gene in Beijing The polymorphism of the receptor (follicle-stimulating hormone receptor, FSHR) gene and the relationship between SNPs and production traits were studied. The expression and expression patterns of FSH beta and FSHR genes in Jinghai yellow chicken were studied by RQ-PCR method. The main results were as follows: 1. the whole exons of the FSH beta gene were detected and the single nucleotide polymorphisms were analyzed. The results showed FS. The H beta gene only exon 3 existed polymorphism, in which the adjacent C2049T and A2050T mutations formed the 3 genotypes of AA, AB and BB; T2341C mutation formed CC, CD and DD 3 genotypes; A2467C mutation formed EE, EE, and 3 genotypes. The correlation analysis showed that the genotype of 12 weeks of age was significantly higher than that of the genotype. EE genotype (P0.05); the 2 week old weight CC genotype was significantly higher than the CD genotype (P0.01), and the above mutation had no significant effect on the reproductive traits. Therefore, the AB, CC and EF genotypes were the dominant genotypes in the growth traits. The FSH beta gene could be preliminarily concluded that the FSH beta gene could promote the growth of Jinghai yellow chicken by.2.. The exons 1 and 10 of the FSHR gene were measured, and the single nucleotide polymorphisms were analyzed. The results showed that there was a C159T mutation in the exon 1 of the FSHR gene, forming three genotypes of AA, AB and BB; 10 of the exons of the FSHR gene had a point mutation of C75429T, forming three genotypes CC, CD and DD. correlation analysis showed that the 2 week age weight AB genotypes of Jinghai yellow chicken were extremely significant. Higher than BB genotype (P0.01), AA genotype was significantly higher than that of BB genotype (P0.05). The 6 week old weight AA and AB genotypes were significantly higher than that of BB genotype (P0.05), and CD genotypes were significantly higher than CC genotype (P0.05). The other differences were not significant.3. on the expression of FSH beta gene in the different growth stages and tissues of Jinghai yellow chicken. The results showed that the expression of FSH beta gene in different tissues had certain expression, the expression level of gonadal and pituitary was the highest, and the expression of FSH beta gene nRNA expression was 20 weeks on the male hen at the age of 0-20 weeks. The expression trend is the same. The relative expression amount is almost zero at the beginning of the pituitary, and there is a small increase in the age of 4 weeks. The expression amount from 4 weeks to 16 weeks old is greatly increased, and the 20 week age of the hen reaches the highest, that is, the expression of FSH beta gene from 4 weeks to 20 weeks is up in a straight line. In the ovary, a small amount of expression is found in the ovary and then 4. In addition, the relative expression of FSH beta gene in the sex glands and pituitary of hens at 4 weeks and 16 weeks old was higher than that of the rooster.4.. The expression of FSHR gene in the different growth stages and tissues of the Beijing yellow chicken was studied. The results showed that the expression of FSHR gene in the sex gland was the highest and the expression abundance was at the beginning of birth. At the age of 0-20 weeks, the expression of FSHR gene in the ovary and testis was decreased at the age of 4 weeks and at the age of 16 weeks. The expression of FSHR gene in the ovary of the 20 week old hen was slightly higher than that at the age of 16 weeks. In the pituitary, the expression of the FSHR gene on the Beijing yellow chicken was slow and smooth. On the other hand, the expression of FSHR gene in the pituitary gland of hens was similar to that in the same period. The expression level of hen's ovaries was slightly higher than that of cock's testis.
【学位授予单位】：扬州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S831

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