授粉后紅球姜雌性生殖器官qRT-PCR的內(nèi)參基因篩選
發(fā)布時間：2019-06-12 04:49
【摘要】：根據(jù)授粉后不同時間點的紅球姜轉(zhuǎn)錄組數(shù)據(jù)庫以及相關(guān)文獻(xiàn)報道的傳統(tǒng)內(nèi)參基因,篩選出10個表達(dá)相對穩(wěn)定的基因Actin-2(ACT2)、Actin-7(ACT7)、Beta tubulin-1(TUB1)、Beta tubulin-5(TUB5)、Alpha tubulin-3(TUA3)、Ubiquitin(UBQ)、Glyceraldehyde-3-phosphate dehydrogenase(GAPDH)、Elongation factor 1-alpha(EF-1α)、Cyclophilin(CYP)、Histone(H2A)作為候選內(nèi)參基因,采用qRT-PCR技術(shù),結(jié)合GeNorm、NormFinder和BestKeeper軟件對候選內(nèi)參基因的表達(dá)穩(wěn)定性進(jìn)行分析.結(jié)果表明,在紅球姜雌性生殖器官授粉后的發(fā)育過程中,GAPDH和UBQ的表達(dá)穩(wěn)定性最好,均適合作為內(nèi)參基因,同時使用2種作為內(nèi)參基因能使實時熒光定量PCR標(biāo)準(zhǔn)化分析結(jié)果更精確.因此,最終選擇GAPDH和UBQ作為實時熒光定量PCR標(biāo)準(zhǔn)化分析紅球姜雌性生殖器官相關(guān)基因表達(dá)的內(nèi)參基因.該研究將為探究紅球姜敗育的分子機(jī)理奠定基礎(chǔ),也為近源姜屬植物內(nèi)參基因的篩選提供線索.
[Abstract]:According to the database of red ginger transcription group and the traditional internal reference genes reported in the literature, 10 relatively stable genes Actin-2 (ACT2) and Actin-7 (ACT7), Beta tubulin-1 (TUB5), Alpha tubulin-3 (TUA3), Ubiquitin (UBQ), Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), Elongation factor 1-alpha (EF-1 偽), Cyclophilin (CYP), Histone (H2A) were selected as candidate internal reference genes. QRT-PCR technique and GeNorm,NormFinder and BestKeeper software were used to analyze the expression stability of candidate internal reference genes. The results showed that the expression stability of GAPDH and UBQ was the best during the development of female reproductive organs of Ginger, and both of them were suitable for internal reference genes. At the same time, the results of real-time fluorescence quantitative PCR standardization analysis could be more accurate by using two kinds of internal reference genes as internal reference genes. Therefore, GAPDH and UBQ were selected as internal reference genes for real-time fluorescence quantitative PCR standardization to analyze the expression of genes related to female reproductive organs of Ginger. This study will lay a foundation for exploring the molecular mechanism of abortion of Ginger, and also provide clues for the screening of internal reference genes in the genus Ginger.
【作者單位】： 華南師范大學(xué)生命科學(xué)學(xué)院廣東省植物發(fā)育與生物工程重點實驗室;
【基金】：國家自然科學(xué)基金委員會-廣東省人民政府聯(lián)合基金(U1301213)
【分類號】：S682.19;Q943.2
本文編號：2497749
[Abstract]:According to the database of red ginger transcription group and the traditional internal reference genes reported in the literature, 10 relatively stable genes Actin-2 (ACT2) and Actin-7 (ACT7), Beta tubulin-1 (TUB5), Alpha tubulin-3 (TUA3), Ubiquitin (UBQ), Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), Elongation factor 1-alpha (EF-1 偽), Cyclophilin (CYP), Histone (H2A) were selected as candidate internal reference genes. QRT-PCR technique and GeNorm,NormFinder and BestKeeper software were used to analyze the expression stability of candidate internal reference genes. The results showed that the expression stability of GAPDH and UBQ was the best during the development of female reproductive organs of Ginger, and both of them were suitable for internal reference genes. At the same time, the results of real-time fluorescence quantitative PCR standardization analysis could be more accurate by using two kinds of internal reference genes as internal reference genes. Therefore, GAPDH and UBQ were selected as internal reference genes for real-time fluorescence quantitative PCR standardization to analyze the expression of genes related to female reproductive organs of Ginger. This study will lay a foundation for exploring the molecular mechanism of abortion of Ginger, and also provide clues for the screening of internal reference genes in the genus Ginger.
【作者單位】： 華南師范大學(xué)生命科學(xué)學(xué)院廣東省植物發(fā)育與生物工程重點實驗室;
【基金】：國家自然科學(xué)基金委員會-廣東省人民政府聯(lián)合基金(U1301213)
【分類號】：S682.19;Q943.2
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