【摘要】：MicroRNAs(miRNAs)大小為20~25個(gè)核苷酸,存在于真核生物體內(nèi),是一類(lèi)內(nèi)源性的具有調(diào)控功能的非編碼RNA。miRNAs通過(guò)堿基互補(bǔ)配對(duì)的方式識(shí)別靶mRNA,并根據(jù)互補(bǔ)程度的不同指導(dǎo)沉默復(fù)合體降解靶mRNA或者阻遏靶mRNA的翻譯。相關(guān)的研究顯示,gga-mir-1662、gga-mir-215和gga-mir-1416與雞腸炎沙門(mén)氏菌的感染相關(guān),且gga-mir-1662與TLR1LA、gga-mir-1416與BCL10可能存在靶向關(guān)系。SNP(single nucleotide polymorphism)是基因組DNA堿基的變異,在基因組中的發(fā)生頻率較高,有意義的變異會(huì)影響到基因組DNA組成元件的功能、轉(zhuǎn)錄產(chǎn)生的mRNA水平、蛋白的表達(dá),進(jìn)而影響基因的功能,導(dǎo)致生物性狀改變甚至致病。因此SNP被廣泛用于群體遺傳學(xué)研究(如生物的起源、進(jìn)化及遷移等方面),和疾病相關(guān)基因的研究,在藥物基因組學(xué)、診斷學(xué)和生物醫(yī)學(xué)研究中起重要作用。本研究從miRNA與靶基因的表達(dá)以及mi RNA的變異兩個(gè)方面研究其在雞腸炎沙門(mén)氏菌感染中的作用機(jī)制。用腸炎沙門(mén)氏菌感染2日齡濟(jì)寧百日雞,在感染后的第1天、第3天、第7天、第14天、第21天、第28天和第35天采集空腸組織樣品,實(shí)驗(yàn)組和對(duì)照組各4只,提取組織總RNA后檢測(cè)相關(guān)microRNA和靶基因的表達(dá)量;感染后第7天取200只濟(jì)寧百日雞的盲腸內(nèi)容物和血液,分別用來(lái)檢測(cè)腸炎沙門(mén)氏菌的含量和提取基因組DNA,設(shè)計(jì)相應(yīng)引物,擴(kuò)增miRNA的目的片段,進(jìn)行測(cè)序、分型,對(duì)SNP位點(diǎn)與腸道內(nèi)容物腸炎沙門(mén)氏菌含量進(jìn)行關(guān)聯(lián)分析。研究結(jié)果顯示:1)不同的microRNA對(duì)照組和實(shí)驗(yàn)組差異表達(dá)的時(shí)間點(diǎn)不同,gga-mir-1662和gga-mir-215在感染后的第7天達(dá)到差異表達(dá)的高峰,試驗(yàn)組的表達(dá)量分別是對(duì)照組的1.6倍和1.66倍。gga-mir-1416則在21日齡表現(xiàn)出明顯的上調(diào)表達(dá),實(shí)驗(yàn)組是對(duì)照組的1.5倍。2)microRNA及其潛在靶基因不是在每個(gè)時(shí)間點(diǎn)能表現(xiàn)出明顯的靶向關(guān)系,可能與機(jī)體的復(fù)雜調(diào)控有關(guān)系。感染后的第7天,gga-mir-1662明顯上調(diào),TLR1LA明顯下調(diào),靶向關(guān)系明顯;gga-mir-1416與BCL10在感染后的第21天和第28天差異顯著,但不能表現(xiàn)出明顯的靶向關(guān)系。3)不同的microRNA在體內(nèi)的變化趨勢(shì)不同,對(duì)照組和實(shí)驗(yàn)組表現(xiàn)出相近的表達(dá)趨勢(shì)。相對(duì)于各個(gè)不同時(shí)間點(diǎn)的表達(dá),總體而言,感染后的第1天、第3天、第7天、第14天屬于高表達(dá)時(shí)間點(diǎn),而第21天、第28天、第35天屬于低表達(dá)時(shí)間點(diǎn)。4)對(duì)個(gè)體分型之后與腸炎沙門(mén)氏菌含量相關(guān)聯(lián),發(fā)現(xiàn)了4個(gè)與腸炎沙門(mén)氏菌感染相關(guān)的SNP位點(diǎn)突變,分別為rs18823870、rs312707374、rs316396584、rs314185118。rs18823870位于gga-mir-215上游,rs312707374、rs316396584、rs314185118位于gga-mir-1416上游,進(jìn)一步說(shuō)明gga-mir-215和gga-mie-1416與腸炎沙門(mén)氏菌感染顯著相關(guān),可以作為抗腸炎沙門(mén)氏菌感染的分子標(biāo)記。本研究鑒定出4個(gè)與腸炎沙門(mén)氏菌菌感染相關(guān)的位于miRNA上游的SNP位點(diǎn),gga-mir-215和gga-mie-1416與腸炎沙門(mén)氏菌感染顯著相關(guān),可以作為抗腸炎沙門(mén)氏菌感染的分子標(biāo)記。
[Abstract]:MicroRNAs (miRNAs) range from 20 to 25 nucleotides and exist in eukaryotes. It is a kind of endogenous non-coding RNA.miRNAs with regulatory function, which recognizes target mRNAs by base complementary pairing, and guides the translation of target mRNA or repressor target mRNA by silencing complex according to the degree of complementarity. Related studies have shown that gga-mir-1662 and TLR1LAGGa-mir-1416 may have a targeting relationship with BCL10, and that gga-mir-1662 and TLR1LAgga-mir-1416 may be a variation of genomic DNA base, and the frequency of occurrence in the genome is higher than that of TLR1LAGgga-mir-1416 and BCL10, and the relationship between gga-mir-1662 and TLR1LAgga-mir-1416 and BCL10 may be related to the infection of Salmonella enteritidis. Meaningful variation will affect the function of the components of genomic DNA, the level of mRNA produced by transcription, the expression of protein, and then affect the function of gene, leading to the change of biological traits and even the pathogenesis of the disease. Therefore, SNP is widely used in population genetics (such as the origin, evolution and migration of organisms) and disease related genes, which play an important role in pharmacogenomics, diagnostics and biomedical research. In this study, we studied the mechanism of miRNA and target gene expression and the variation of miRNA in salmonella enteritidis infection. Two days old Jining 100 day chickens infected with Salmonella enteritidis were collected on the 1st, 3rd, 7th, 14th, 21st, 28th and 35th day after infection. The cecum contents and blood of 200 Jining 100 day chickens were collected on the 7th day after infection to detect the content of Salmonella enteritidis and to extract the genomic DNA, and to design the corresponding primer, which were used to detect the content of Salmonella enteritidis and to extract the genomic DNA. The aim fragment of miRNA was amplified, sequenced and typed, and the correlation between SNP site and intestinal content of Salmonella enteritis was analyzed. The results showed that the different time points of different microRNA expression in control group and experimental group reached the peak of differential expression on the 7th day after infection. The expression levels of microRNA and its potential target genes in the experimental group were 1.6 times and 1.66 times as much as those in the control group, respectively, and were significantly up-regulated at 21 days of age, while those in the experimental group were 1.5 times higher than those in the control group. It may be related to the complex regulation of the body. On the 7th day after infection, TLR1LA was up-regulated and down-regulated, and the targeting relationship was significantly different between BCL10 and BCL10 on the 21st and 28th days after infection, but there was no obvious targeting relationship between BCL10 and BCL10 on the 21st and 28th days after infection. The control group and the experimental group showed similar expression trend. Compared with the expression at different time points, on the whole, the first day, the third day, the seventh day, the 14th day after infection were high expression time points, and the 21st day, the 28th day, On the 35th day, four SNPs related to salmonella enteritis were found, which were rs18823870rs312707374rs316396584rs316396584rs1838707374rs1838707374rs3127374rs312396584rs312707374rs312396584rs312707374rs316396584rs31485118 in the upper reaches of gga-mir-215. The results further indicated that gga-mir-215 and gga-mie-1416 were significantly correlated with Salmonella enteritis infection and could be used as molecular markers against Salmonella enteritidis infection. In this study, four SNPs located upstream of gga-mie-1416 and SNPs related to the infection of Salmonella enteritidis were identified, which could be used as molecular markers against Salmonella enteritidis infection.
【學(xué)位授予單位】：山東農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：S858.31

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 Li Ma;Mei-Sze Chua;Ourania Andrisani;Samuel So;;Epigenetics in hepatocellular carcinoma: An update and future therapy perspectives[J];World Journal of Gastroenterology;2014年02期

2 唐一通;肖娜;李智山;范曉航;余燕敏;姚勁松;;單核苷酸多態(tài)性檢測(cè)方法研究進(jìn)展[J];現(xiàn)代生物醫(yī)學(xué)進(jìn)展;2013年27期

3 張哲;張勤;丁向東;;畜禽基因組選擇研究進(jìn)展[J];科學(xué)通報(bào);2011年26期

4 孔繁飛;孫朝陽(yáng);王中顯;韓凌斐;翁丹卉;盧運(yùn)萍;陳剛;;miR-125b Confers Resistance of Ovarian Cancer Cells to Cisplatin by Targeting Pro-apoptotic Bcl-2 Antagonist Killer 1[J];Journal of Huazhong University of Science and Technology(Medical Sciences);2011年04期

5 韓建文;張學(xué)軍;;全基因組關(guān)聯(lián)研究現(xiàn)狀[J];遺傳;2011年01期

6 張哲;許德義;蔣曉飛;;變性梯度凝膠電泳檢測(cè)鮑曼不動(dòng)桿菌中SHV基因單核苷酸多態(tài)性[J];現(xiàn)代實(shí)用醫(yī)學(xué);2009年08期

7 郭闖;高原;王煒;;動(dòng)物性食品中沙門(mén)氏菌檢測(cè)技術(shù)研究進(jìn)展[J];內(nèi)蒙古民族大學(xué)學(xué)報(bào)(自然科學(xué)版);2008年03期

8 Soo Ryang Kim;Hirotsugu Ikawa;Kenji Ando;Keiji Mita;Shuichi Fuki;Michiie Sakamoto;Yoshihiro Kanbara;Toshiyuki Matsuoka;Masatoshi Kudo;Yoshitake Hayashi;;Multistep hepatocarcinogenesis from a dysplastic nodule to well-differentiated hepatocellular carcinoma in a patient with alcohol-related liver cirrhosis[J];World Journal of Gastroenterology;2007年08期

9 RN Zárate R;F Arias;E Bandres;E Cubedo;R Malumbres;J García-Foncillas;;Xeroderma pigmentosum group D 751 polymorphism as a predictive factor in resected gastric cancer treated with chemo-radiotherapy[J];World Journal of Gastroenterology;2006年37期

10 魏笑笑;王寶維;荊麗珍;;家禽抗病育種研究進(jìn)展[J];家禽科學(xué);2006年07期

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