本文選題：民豬 + 外周血單核細(xì)胞　； 參考：《中國(guó)畜牧獸醫(yī)》2017年12期

【摘要】：民豬是能夠適應(yīng)東北地區(qū)寒冷環(huán)境的地方豬種,但目前對(duì)其環(huán)境適應(yīng)性相關(guān)基因的研究較少,也沒有確定適合此研究所需的實(shí)時(shí)熒光定量PCR的內(nèi)參基因。因此,本試驗(yàn)通過研究常用的12個(gè)內(nèi)參基因(B2M、ACTB、RPL11、RPL4、YWHAZ、GAPDH、HPRT1、SDHA、HMBS、IDH3B、TUBB2B和TBP1)在不同環(huán)境溫度下民豬外周血單核細(xì)胞中的表達(dá)穩(wěn)定性,旨在確定合適的內(nèi)參基因進(jìn)行相關(guān)研究。分別在-25、5、10和30℃采集3頭民豬耳靜脈血分離出單核細(xì)胞,利用geNorm、NormFinder、BestKeeper 3種軟件分析12個(gè)候選內(nèi)參基因的Ct值,篩選出表達(dá)穩(wěn)定的基因作為內(nèi)參基因。經(jīng)geNorm和NormFinder計(jì)算獲得各候選基因的M值發(fā)現(xiàn),12個(gè)候選基因的表達(dá)均相對(duì)穩(wěn)定,而BestKeeper的分析則顯示ACTB、GAPDH、SDHA、HPRT1、TBP1、YWHAZ基因的SD值均1,可用作本研究條件下的內(nèi)參基因,而另外6個(gè)基因SD值則1,不符合作為內(nèi)參基因的標(biāo)準(zhǔn)。綜合3種分析的結(jié)果,在本研究條件下,TBP1基因的穩(wěn)定性最高,ACTB、GAPDH、SDHA、HPRT1和YWHAZ基因也符合作為內(nèi)參基因的標(biāo)準(zhǔn),都可研究在不同環(huán)境溫度下民豬外周血單核細(xì)胞中基因表達(dá)時(shí)作為內(nèi)參基因。
[Abstract]:Minzhou pig is a local pig that can adapt to cold environment in Northeast China, but there are few studies on the genes related to its environmental adaptability, and there is no internal reference gene for real-time fluorescent quantitative PCR which is suitable for this study. Therefore, in this experiment, the expression stability of 12 common domestic reference genes (B2MN, ACTBN, RPL11, RPL4, YWHAZN, GAPDH1, HPRT1) in human peripheral blood mononuclear cells (PBMC) was studied in order to determine the appropriate endogenous genes in human peripheral blood mononuclear cells (PBMC) at different ambient temperatures, so as to determine the expression stability of HMBSIDH3BUBB2B and TBP1 in human peripheral blood mononuclear cells (PBMC). Monocytes were isolated from venous blood of three human pig ears at -25 鈩，

本文編號(hào)：1863866