發(fā)布時間：2018-04-13 04:26

  本文選題：中國美利奴綿羊 + CCNY基因��； 參考：《畜牧獸醫(yī)學(xué)報》2017年08期

【摘要】：旨在驗證Cyclin Y(CCNY)基因與羊毛品質(zhì)性狀的相關(guān)性,并探索其可能的作用機(jī)制。本研究以744只中國美利奴羊(新疆軍墾型)為試驗材料,采用Sanger法混池測序?qū)ふ叶鄳B(tài)性位點;利用MALDI-TOF-MS方法進(jìn)行個體基因型鑒定;利用real-time RT-PCR檢測基因表達(dá);利用免疫熒光染色技術(shù)分析蛋白定位;利用CCK-8方法檢測細(xì)胞的增殖;利用熒光素酶報告基因技術(shù)檢測Wnt/β-catenin信號通路的活性。結(jié)果表明,本研究鑒定出中國美利奴羊CCNY基因的3個SNPs位點,分別命名為CCNYSNP1(rs613860955)、CCNYSNP2(rs622396422)和CCNYSNP3(rs611417387)。關(guān)聯(lián)分析顯示,這3個SNPs及其構(gòu)建的單倍型均與羊毛細(xì)度顯著相關(guān)(P0.05);組織表達(dá)譜檢測分析顯示,CCNY基因在睪丸和肺中有較高的表達(dá),在皮膚等組織中有中等程度的表達(dá);免疫熒光分析顯示,CCNY主要定位于毛囊的內(nèi)根鞘和毛母質(zhì)細(xì)胞;CCK-8細(xì)胞增殖檢測分析顯示,過表達(dá)CCNY基因顯著促進(jìn)人表皮永生化細(xì)胞(HaCaT)和綿羊胎兒成纖維細(xì)胞(SFF)的增殖(P0.05);報告基因分析結(jié)果顯示,過表達(dá)CCNY基因極顯著促進(jìn)Wnt/β-catenin信號通路的活性(P0.01)。以上結(jié)果顯示,CCNY是控制羊毛細(xì)度的一個重要基因,所鑒定出的CCNY基因3個SNPs有望用于細(xì)毛羊的分子輔助選育。
[Abstract]:The aim of this study was to verify the correlation between Cyclin YC gene and wool quality traits, and to explore its possible mechanism of action.In this study, 744 Chinese Merino sheep (Xinjiang Army Reclamation) were used to search for polymorphic loci by Sanger mixed pool sequencing, individual genotypes were identified by MALDI-TOF-MS method, and gene expression was detected by real-time RT-PCR.The protein localization was analyzed by immunofluorescence staining, the proliferation of cells was detected by CCK-8 method, and the activity of Wnt/ 尾 -catenin signaling pathway was detected by luciferase reporter gene technique.The results showed that three SNPs loci of the CCNY gene of Chinese Merino sheep were identified and named as CCNYSNP1 rs613860955) and CCNYSNP3 rs611417387 respectively.Correlation analysis showed that the three SNPs haplotypes and their haplotypes were significantly correlated with wool fineness (P0.05A), and the expression of CCNY gene was higher in testis and lungs and moderately expressed in skin and other tissues.Immunofluorescence analysis showed that CCNY was mainly located in the inner root sheath of hair follicles and the proliferation of CCK-8 cells in hair follicles.Overexpression of CCNY gene significantly promoted the proliferation of human epidermal immortalized cells (HaCaT) and sheep fetal fibroblast (SFF), and the results of report gene analysis showed that overexpression of CCNY gene significantly promoted the activity of Wnt/ 尾 -catenin signaling pathway.The results showed that CCNY was an important gene to control wool fineness, and the three SNPs genes identified in this study were expected to be used in molecular assisted selection of fine wool sheep.
【作者單位】： 農(nóng)業(yè)部雞遺傳育種重點實驗室;黑龍江省動物細(xì)胞與遺傳工程重點實驗室;新疆農(nóng)墾科學(xué)院畜牧獸醫(yī)研究所;
【基金】：國家轉(zhuǎn)基因?qū)ｍ椪n題(2014ZX08009-002;2009ZX08009-160B)
【分類號】：S826.2

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