本文選題：東北虎 + 肝臟�。� 參考：《東北林業(yè)大學(xué)》2016年碩士論文

【摘要】：東北虎(Panthera tigris altaica)是國(guó)家Ⅰ級(jí)重點(diǎn)保護(hù)動(dòng)物,也是全球最瀕危的物種之一。長(zhǎng)期以來,東北虎生存環(huán)境的嚴(yán)重破壞,獵物資源匱乏,人類亂捕亂殺因素,導(dǎo)致野生東北虎處于絕滅的境地,及時(shí)開展東北虎遺傳資源保存及相關(guān)分子生物學(xué)研究,具有非常重要科學(xué)價(jià)值和保護(hù)意義。本文選取圈養(yǎng)東北虎的肝臟為研究材料,采用現(xiàn)代分子生物學(xué)的實(shí)驗(yàn)方法,構(gòu)建了東北虎肝臟的cDNA文庫(kù),同時(shí)對(duì)該cDNA文庫(kù)的質(zhì)量加以分析鑒定,獲得出如下的研究結(jié)果：1、以2.5歲齡的東北虎胎肝臟組織為研究材料,獲得了高質(zhì)量東北虎胎盤cDNA文庫(kù)。原始文庫(kù)庫(kù)容量為2.68×106pfu/ml,重組率為97.2%,擴(kuò)增后的文庫(kù)庫(kù)容量達(dá)到4.8×109pfu/ml,重組率為94.6%,隨機(jī)挑選200個(gè)陽性菌斑進(jìn)行PCR檢測(cè),獲得的文庫(kù)中插入片段的長(zhǎng)度范圍在0.5～3.0Kb之間,并且片段的平均長(zhǎng)度為0.78Kb。2、通過隨機(jī)挑選200個(gè)克隆展開測(cè)序工作,運(yùn)用GENE BANK中的BLAST對(duì)測(cè)得的序列進(jìn)行比對(duì)分析,獲得了138個(gè)原始序列,去除掉測(cè)序質(zhì)量比較差和很短(200bp)的序列,最后獲得了105條有效的ESTs序列。大部分序列長(zhǎng)度都大于300bp,主要集中的在600～900bp之間,測(cè)序成功率達(dá)到89.3%。這些序列經(jīng)過GO分析分成三大類,即細(xì)胞組成,分子功能,生物進(jìn)程。3、在GO分類結(jié)果中,80個(gè)序列中,有71個(gè)被注釋,10種分子功能被記錄了107次,有12個(gè)參與的生物學(xué)過程以及6種細(xì)胞組分。GO分類發(fā)現(xiàn)同基因有不同的分子功能,也能參與不同的生物學(xué)過程,這反映了東北虎生命過程的復(fù)雜性。4、將80個(gè)基因進(jìn)行蛋白質(zhì)分類及代謝通路分析,得到24種蛋白質(zhì)及33個(gè)信號(hào)通路。并對(duì)重要的蛋白質(zhì)及代謝通路進(jìn)行分析。5、通過構(gòu)建肝組織相關(guān)基因相互作用的網(wǎng)絡(luò)圖,并運(yùn)用GO注釋,KEGG代謝途徑分析和InterPro檢索等工具以及對(duì)相關(guān)文獻(xiàn)的查找,篩選出肝組織中相關(guān)的基因,例如SOD1,CTNNB1,LIF2等,對(duì)這些基因的功能進(jìn)行研究,從而能夠提高對(duì)于東北虎新陳代謝的生物學(xué)角度的認(rèn)知程度,增強(qiáng)東北虎自身體質(zhì)并且提高其生存質(zhì)量。
[Abstract]:Panthera tigris altaica) is one of the most endangered species in the world. For a long time, the serious destruction of the living environment of the Siberian tiger, the scarcity of prey resources, and the human random hunting and killing factors have led to the extinction of the wild Siberian tiger. The preservation of the genetic resources of the Siberian tiger and the related molecular biology research have been carried out in time. It has very important scientific value and protective significance. In this paper, the liver of the captive Siberian tiger was selected as the research material, and the cDNA library of the liver of the Siberian tiger was constructed by using the modern molecular biology method, and the quality of the cDNA library was analyzed and identified. The following results were obtained: 1: 1. A high quality placental cDNA library of Amur tiger was obtained from the liver tissue of a 2.5 year old Siberian tiger fetus. The original library capacity was 2.68 脳 106pfuP / ml, the recombination rate was 97.2ml, the expanded library capacity reached 4.8 脳 109pfuP / ml, and the recombination rate was 94.6. 200 positive plaque were randomly selected for PCR detection, and the length of inserted fragments in the library was between 0.5~3.0Kb. And the average length of the fragment is 0.78 Kb. 2. By randomly selecting 200 clones to carry out the sequencing work, using BLAST in GENE BANK to analyze the measured sequence, 138 original sequences were obtained, and the sequence of poor quality and very short 200bp was removed. Finally, 105 valid ESTs sequences are obtained. Most of the sequences were longer than 300 BP, mainly between 600~900bp and the success rate of sequencing was 89.33%. These sequences are classified into three categories by go analysis: cell composition, molecular function, biological process. Out of 80 sequences, 71 have been annotated for 10 molecular functions 107 times. There are 12 involved biological processes and 6 cell fractions. Go classification shows that the same gene has different molecular functions and can also participate in different biological processes. This reflects the complexity of the life process of Amur tiger. The protein classification and metabolic pathway analysis of 80 genes obtained 24 proteins and 33 signal pathways. The important proteins and metabolic pathways were analyzed. 5. By constructing a network map of the interaction of genes related to liver tissue, and using go annotated KEGG metabolic pathway analysis and InterPro search tools, and searching the relevant literature, and so on. The related genes in liver tissue, such as SOD1 CTNNB1LIF2, were screened, and the function of these genes was studied, so as to improve the cognition of the biological angle of Amur tiger metabolism, enhance the physical constitution and improve the quality of life of Amur tiger.
【學(xué)位授予單位】：東北林業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：Q953

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