本文選題：棉酚　切入點：IFN-γ　出處：《塔里木大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：棉酚，又名棉毒素或者棉籽餅，是棉屬植物內(nèi)形成的一種多酚類化合物存在于植株各部分的褐色點狀色素腺體中，而棉籽餅粕確是一種產(chǎn)量大，價格低廉，富含蛋白質(zhì)的動物飼料，可就是因為棉酚的毒性限制了這種優(yōu)良飼料的大規(guī)模的應(yīng)用。無論是對反芻動物還是非反芻動物都有毒性，且呈劑量依賴性，，會對動物的大腦、心、肝、血管等器官造成損害，致使家畜流產(chǎn)。IFN-γ是由免疫細胞分泌的一種細胞因子，是一種重要的免疫調(diào)節(jié)因子，能調(diào)節(jié)和激活T淋巴細胞的活性，以應(yīng)對外界抗原的侵入，維持機體的平衡。為了探討棉酚對免疫系統(tǒng)的影響以及IFN-γ在棉酚作用于免疫系統(tǒng)中的作用，本實驗以新疆多浪羊為實驗對象，分離多浪羊的外周血淋巴細胞，通過不同濃度的棉酚與淋巴細胞體外共培養(yǎng)，利用熒光定量PCR的方法，檢測IFN-γ的表達量的變化，結(jié)果，隨著棉酚濃度的不斷遞增，IFN-γ的表達量也呈逐漸增長的趨勢。 利用PCR的方法擴增IFN-γ的CDS區(qū)，通過HindIII和BamHI對pMD18T-IFN-γ和pcDNA3.1進行雙酶切，連接，轉(zhuǎn)化，測序和序列分析，結(jié)果成功克隆出IFN-γ的CDS區(qū)，序列長度455bp，比對結(jié)果表明其與羊的IFN-γ序列的一致性達到99%以上。進一步分離外周血淋巴細胞，利用RPMI1640完全培養(yǎng)基體外培養(yǎng)淋巴細胞，待細胞進入對數(shù)期，利用電轉(zhuǎn)染系統(tǒng)，將pcDNA3.1-IFN-γ真核表達載體轉(zhuǎn)入淋巴細胞中，利用半定量的方法檢測轉(zhuǎn)染組與對照組的IFN-γ的表達量。結(jié)果表明，通過比較轉(zhuǎn)染組和對照組IFN-γ的表達量，轉(zhuǎn)染組明顯高于對照組。
[Abstract]:Gossypol, also known as cotton toxin or cottonseed cake, is a polyphenolic compound formed in cotton plants. Protein-rich animal feed, however, is limited by the toxicity of gossypol to the large-scale use of this fine feed. Both ruminant and non-ruminant animals are toxic, and in a dose-dependent manner, it affects the brains and hearts of animals. The liver, blood vessels and other organs are damaged, causing livestock abortion. IFN- 緯 is a cytokine secreted by immune cells and is an important immunomodulating factor, which can regulate and activate the activity of T lymphocytes in response to the invasion of external antigens. In order to study the effect of gossypol on immune system and the effect of IFN- 緯 on immune system, the peripheral blood lymphocytes of Doulang sheep in Xinjiang were isolated. By co-culture of gossypol and lymphocytes in vitro, the expression of IFN- 緯 was detected by fluorescence quantitative PCR. The results showed that the expression of IFN- 緯 increased with the increasing of gossypol concentration. The CDS region of IFN- 緯 was amplified by PCR. PMD18T-IFN- 緯 and pcDNA3.1 were digested, ligated, transformed, sequenced and sequenced by HindIII and BamHI. The CDS region of IFN- 緯 was cloned successfully. The sequence length was 455bp. the result of alignment showed that the consistency of IFN- 緯 sequence with sheep was more than 99%. The peripheral blood lymphocytes were further isolated and cultured in vitro on RPMI1640 complete medium, the cells were in logarithmic phase, and the electrotransfection system was used. The eukaryotic expression vector pcDNA3.1-IFN- 緯 was transferred into lymphocytes, and the expression of IFN- 緯 in transfection group and control group was detected by semi-quantitative method. The results showed that the expression of IFN- 緯 in transfection group was significantly higher than that in control group by comparing the expression of IFN- 緯 between transfection group and control group.
【學(xué)位授予單位】：塔里木大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S826.5

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