本文選題：綿羊　切入點：胎兒　出處：《中國農(nóng)業(yè)科學院》2015年碩士論文

【摘要】：綿羊是重要的經(jīng)濟動物,肉用性狀選育已成為綿羊育種的主要方向。不同品種綿羊生長調(diào)控機制存在差異,對其開展研究有助于發(fā)現(xiàn)未知基因調(diào)控作用。Lnc RNA是一類長度大于200nt的非編碼RNA分子,在哺乳動物肌肉發(fā)育具有重要調(diào)控作用,且時空差異表達明顯。本研究選用發(fā)育明顯差異的兩品種烏珠穆沁和特克賽爾綿羊,利用二代測序技術分別對妊娠85d、120d、初生和一周歲腓腸肌進行分析,從lnc RNA方面揭示兩品種肉用性能差異的內(nèi)在機制。同時在細胞水平對篩選到的TCONS_00102859進行試驗分析。主要試驗結果如下:1.綿羊lnc RNA測序及描述型分析測序數(shù)據(jù)經(jīng)篩選處理后平均每個樣本獲得2.25 Gb數(shù)據(jù),其中85日齡胎兒數(shù)據(jù)量最小,過濾數(shù)據(jù)質(zhì)量可靠。過濾數(shù)據(jù)回帖基因組,整體比對率≥89%,組裝后進行l(wèi)nc RNA篩選,共獲得2002條候選lnc RNA。Lnc RNA在綿羊染色體上不存在偏好性,長度跨越大,但78.5%的lnc RNA長度2000nt。候選lnc RNA平均外顯子數(shù)目為2.8個,90%的lnc RNA外顯子數(shù)少于4個。比對后發(fā)現(xiàn)1967條為新獲得轉錄本。2.候選lnc RNA靶基因注釋靶基因GO注釋分析發(fā)現(xiàn),篩選到的lnc RNA可能與結合和催化作用、代謝過程、色素沉著和繁殖性能等相關。KEGG注釋分析發(fā)現(xiàn),信號轉導、易感疾病、癌癥、轉錄和翻譯等通路注釋到最多基因。3.特克賽爾和烏珠穆沁相同時期差異lnc RNA分析不同品種相同發(fā)育階段差異lnc RNA分析發(fā)現(xiàn),每個特定階段兩個品種差異表達lnc RNA靶基因富集結果有明顯差異。85日齡胎兒富集到MAPK信號通路、Wnt信號通路等胚胎肌肉發(fā)育、腫瘤發(fā)生和其他相關通路;120日齡階段富集到心臟機能相關通路;初生階段富集到磷脂酰肌醇信號系統(tǒng)和鈣信號等通路;成年階段富集到煙酸和煙酰胺代謝通路。所有發(fā)育階段兩個品種都有差異表達的lnc RNA篩選到10個。其中有三個lnc RNA,TCONS_00051148、TCONS_00079077和TCONS_00078553,對應到基因內(nèi)部,對應的基因分別為ANKRD10、FGF2和LOC101105780;TCONS_00102859只在特克賽爾表達且表達量逐漸升高。4.TCONS_00102859在綿羊成肌細胞功能初步分析構建TCONS_00102859慢病毒過表達載體,在綿羊原代成肌細胞上初步驗證其作用。發(fā)現(xiàn)GV287-TCONS_00102859對原代成肌細胞具有較高的轉染效率,且可以顯著提高TCONS_00102859在細胞內(nèi)表達量。轉染細胞中Myo D、Myo G和Mef2c表達上調(diào),MSTN表達下調(diào),初步證明TCONS_00102859可能對成肌細胞的增殖和分化均有一定作用。本研究進行的綿羊lnc RNA轉錄組的特征分析與細胞水平功能初步分析,將進一步促進綿羊lnc RNA功能和基因調(diào)控機制的研究。
[Abstract]:Sheep is an important economic animal and breeding of meat traits has become the main direction of sheep breeding. It is helpful to find that the unknown gene regulation. LNC RNA is a class of noncoding RNA molecules which are longer than 200nt and play an important role in the regulation of mammalian muscle development. In this study, two different breeds of sheep, Wuzhimuqin and Texer, were used to analyze the gastrocnemius muscle of 85 days gestation at 120 days after pregnancy, using the second generation sequencing technique. The internal mechanism of meat performance difference between the two breeds was revealed from the aspect of lnc RNA. At the same time, the selected TCONS_00102859 was tested and analyzed at the cell level. The main results were as follows: 1.The sequencing data of sheep lnc RNA and descriptive analysis were screened. After processing, an average of 2.25 GB data per sample was obtained, Among them, 85 day-old fetuses had the smallest amount of data and the quality of filtered data was reliable. After filtering the genome, the overall comparison rate was 鈮，

本文編號：1695717