發(fā)布時(shí)間：2018-03-10 01:05

  本文選題：鹿茸　切入點(diǎn)：干細(xì)胞　出處：《農(nóng)業(yè)生物技術(shù)學(xué)報(bào)》2017年05期 　論文類型：期刊論文

【摘要】：合適的內(nèi)參基因是利用qRT-PCR準(zhǔn)確分析基因相對(duì)表達(dá)量的先決條件。為了對(duì)鹿茸生長(zhǎng)中心細(xì)胞和鹿茸干細(xì)胞間的差異表達(dá)基因進(jìn)行準(zhǔn)確定量分析,本研究篩選了TATA盒結(jié)合蛋白(TATA box binding protein,TBP)、β-肌動(dòng)蛋白(actin beta,ACTB)、微管蛋白1α(tubulin alpha 1a,TUBA1A)、葡萄糖-6-磷酸脫氫酶(glucose-6-phosphate dehydrogenase,G6PDH)、3-磷酸甘油醛脫氫酶(glyceraldehyde-3-phosphate dehydrogenase,GAPDH)及β2-微球蛋白(β2-microglobulin,B2M)這6個(gè)基因作為內(nèi)參基因,利用qRT-PCR技術(shù)分別檢測(cè)這些基因m RNA在鹿茸生長(zhǎng)中心細(xì)胞及鹿茸干細(xì)胞中的表達(dá)情況,并對(duì)其表達(dá)的穩(wěn)定性進(jìn)行了綜合評(píng)估。運(yùn)用Delta-Ct method,ge Norm(ver.3.5)、Bestkeeper(ver.1.0)和NormFinder(ver.0.953)和Ref Finder軟件綜合分析表明,在鹿茸干細(xì)胞之間,ACTB和TUBA1A基因表達(dá)穩(wěn)定性最高;在鹿茸生長(zhǎng)中心細(xì)胞之間,B2M和ACTB基因表達(dá)穩(wěn)定性最高;在所有鹿細(xì)胞系之間,B2M和ACTB基因表達(dá)穩(wěn)定性最高。因此,B2M、ACTB以及TUBA1A候選基因可作為研究鹿茸生長(zhǎng)中心細(xì)胞及鹿茸干細(xì)胞基因差異表達(dá)的內(nèi)參基因,本研究為進(jìn)一步研究鹿茸生長(zhǎng)發(fā)育機(jī)制和開(kāi)展鹿茸模型應(yīng)用于生物醫(yī)學(xué)研究提供了一定的理論依據(jù)。
[Abstract]:In order to analyze the differentially expressed genes between antler growth center cells and pilose antler stem cells accurately, it is a prerequisite for accurate analysis of relative expression by qRT-PCR. In this study, Tata box binding TATA cassette binding protein, 尾 -actin betaan ACTB, tubulin 1 偽 tubulin alpha TUBA1A, glucose-6-phosphate dehydrogenase G6PDH3-phosphate glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and 尾 2-microglobulinB2M (尾 2-microglobulinB2M) were selected as internal reference genes. QRT-PCR technique was used to detect the expression of m RNA in antler growth center cells and pilose antler stem cells, and the stability of the expression was evaluated. The results of Delta-Ct methodge Normver.3.5Bestkeeperver.1.0) and Norm Finderver.0.953) and Ref Finder software showed that, The stability of ACTB and TUBA1A gene expression was the highest in the pilose antler stem cells, and the stability of B2M and ACTB gene expression was the highest in the pilose antler growth center cells. The stability of B2M and ACTB gene expression was the highest among all deer cell lines. Therefore, the candidate genes of B2M ACTB and TUBA1A could be used as internal reference genes to study the differentially expressed genes of pilose antler growth center cells and pilose antler stem cells. This study provides a theoretical basis for further study on the growth and development mechanism of velvet antler and the application of antler model in biomedical research.
【作者單位】： 中國(guó)農(nóng)業(yè)科學(xué)院特產(chǎn)研究所特種經(jīng)濟(jì)動(dòng)物分子生物學(xué)國(guó)家重點(diǎn)實(shí)驗(yàn)室/吉林省鹿茸工程研究中心;
【基金】：國(guó)家自然科學(xué)基金(No.31500792和No.31402059) 中國(guó)農(nóng)業(yè)科學(xué)院科技創(chuàng)新工程項(xiàng)目(No.CAAS-ASTIP-2014-ISAPS-04)
【分類號(hào)】：S825
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本文編號(hào)：1591060