【摘要】：鵝多數(shù)品種具有明顯的季節(jié)性繁殖與及強(qiáng)烈的就巢性等特點(diǎn),嚴(yán)重制約了鵝業(yè)發(fā)展。籽鵝是世界高產(chǎn)蛋鵝品種之一,因其體型小、產(chǎn)蛋多等特點(diǎn),素有“產(chǎn)蛋王”之美稱。家禽屬于卵生動(dòng)物,在適宜環(huán)境條件下,卵巢上的卵泡可持續(xù)發(fā)育、成熟、排卵,發(fā)育到性成熟的時(shí)候受精,且家禽排卵后不會(huì)有黃體形成,是較為理想的動(dòng)物試驗(yàn)?zāi)Ｐ�。本試�?yàn)以籽鵝作為試驗(yàn)動(dòng)物,圍繞鵝等級(jí)前卵泡發(fā)育調(diào)控機(jī)制展開(kāi)研究。神經(jīng)營(yíng)養(yǎng)因子家族((Neurotrophic factors,NTFs)屬于蛋白質(zhì)類大分子,由多種神經(jīng)營(yíng)養(yǎng)因子組成,其中腦源性神經(jīng)營(yíng)養(yǎng)因子(Brain-derived neurotrophie factor,BDNF))、神經(jīng)生長(zhǎng)因子(Nerve growth factor.NGF),神經(jīng)營(yíng)養(yǎng)因子-3(Neurotrophin-3)是該家族中的重要成員,同時(shí)也是發(fā)現(xiàn)最早、當(dāng)前研究較為熱門的神經(jīng)營(yíng)養(yǎng)因子�，F(xiàn)有研究表明,NGF、NT-3以及BDNF與其受體TrkB基因不僅對(duì)神經(jīng)細(xì)胞具有調(diào)控作用,在心血管、內(nèi)分泌、免疫以及繁殖等系統(tǒng)中也有相應(yīng)的表達(dá)。在鵝卵泡發(fā)育過(guò)程中神經(jīng)營(yíng)養(yǎng)因子有關(guān)基因及其代謝產(chǎn)物起著重要作用,為了解神經(jīng)營(yíng)養(yǎng)因子以及具有調(diào)控作用基因的生物學(xué)信息以及相關(guān)基因的表達(dá)趨勢(shì),本研究采用高通量轉(zhuǎn)錄組測(cè)序技術(shù)。它是組織或細(xì)胞不同生理狀態(tài)下轉(zhuǎn)錄出來(lái)的所有RNA的總和。本試驗(yàn)主要以籽鵝等級(jí)前卵泡作為研究對(duì)象,選用新一代Illumina Hiseq 2500高通量測(cè)序平臺(tái)對(duì)提取的籽鵝等級(jí)前卵泡(5個(gè)等級(jí))的總RNA進(jìn)行轉(zhuǎn)錄組測(cè)序。本次轉(zhuǎn)錄組測(cè)序,共得到長(zhǎng)度在150 bp左右的reads上百萬(wàn)條。鵝歸屬非模式生物,尚未有參考基因組,通過(guò)對(duì)鵝等級(jí)前卵泡優(yōu)質(zhì)序列的拼接組裝以及GO、COG數(shù)據(jù)庫(kù)注釋,挖掘出與繁殖、代謝、生長(zhǎng)速度甚至抗病等相關(guān)的候選基因,為以后實(shí)踐分子育種以及功能基因研究提供理論基礎(chǔ);KEGG pathway注釋到ERK信號(hào)通路以及該通路上的主要基因,計(jì)算FPKM值作為基因的表達(dá)量,利用RT-PCR和Western Blot技術(shù),分析鵝等級(jí)前卵泡中具有主要調(diào)控作用的NT3、NGF、BDNF及其受體TrkB基因的表達(dá),結(jié)果如下:4個(gè)基因在鵝5個(gè)等級(jí)前卵泡中均有表達(dá),其中在小白卵泡中的表達(dá)量最多。
[Abstract]:Most goose varieties have obvious characteristics of seasonal reproduction and strong nesting, which seriously restrict the development of goose industry. Seed goose is one of the high-yielding egg geese in the world. Poultry belongs to oviparous animal. Under suitable environment, follicle on ovary can develop continuously, mature, ovate, fertilize when developing to sexual maturity, and there will not be luteal body formation after ovulation of poultry. It is an ideal animal experimental model. In this study, the control mechanism of prefollicular development of geese was studied with Seedling Goose as experimental animal. The family of neurotrophic factors (Neurotrophic factors,NTFs) belongs to protein macromolecules and consists of a variety of neurotrophic factors, including Brain-derived neurotrophie factor,BDNF), neurotrophic factor (Brain-derived neurotrophie factor,BDNF), neurotrophic factor (Nerve growth factor.NGF), and neurotrophic factor (Nerve growth factor.NGF). Neurotrophic factor-3 (Neurotrophin-3) is an important member of the family and is the most popular neurotrophic factor. Current studies have shown that NGF,NT-3, BDNF and its receptor TrkB genes not only regulate nerve cells, but also express them in cardiovascular, endocrine, immune and reproductive systems. Neurotrophin-related genes and their metabolites play an important role in the development of goose follicles. In order to understand the biological information of neurotrophic factors and regulatory genes and the expression trend of related genes, neurotrophic factors and their related genes play an important role in the development of goose follicles. High-throughput transcriptome sequencing was used in this study. It is the sum of all RNA transcribed by tissues or cells in different physiological states. A new generation of Illumina Hiseq 2500 high throughput sequencing platform was used to sequence the total RNA of prefollicles (5 grades) extracted from the seed geese. The transcriptome was sequenced and millions of reads about 150 bp in length were obtained. Geese belong to non-model organisms without reference genomes. Candidate genes related to reproduction, metabolism, growth rate and even disease resistance are mined by splicing and assembling high-quality pre-follicular sequences and annotating GO,COG database. It provides a theoretical basis for the practice of molecular breeding and functional gene research in the future. KEGG pathway annotates the ERK signaling pathway and the major genes in the pathway, calculates the FPKM value as the expression amount of the gene, and uses RT-PCR and Western Blot techniques to analyze the NT3,NGF, which plays a major role in the pre-grade follicles of goose. The expression of BDNF and its receptor TrkB gene was as follows: the four genes were expressed in goose prefollicles of 5 grades, and most of them were expressed in small white follicles.
【學(xué)位授予單位】：吉林農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S835

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本文編號(hào)：2359738