本文選題：羊泰勒蟲(chóng)　切入點(diǎn)：蛋白組OPRT　出處：《中國(guó)農(nóng)業(yè)科學(xué)院》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：羊泰勒蟲(chóng)病是由媒介蜱傳播的泰勒科泰勒屬的原蟲(chóng)寄生于綿羊和山羊的巨噬細(xì)胞、淋巴細(xì)胞和紅細(xì)胞內(nèi)而引起的疾病的總稱,它給全球養(yǎng)羊業(yè)帶來(lái)了巨大的經(jīng)濟(jì)損失。本研究運(yùn)用生物信息學(xué)初步分析了羊泰勒蟲(chóng)蛋白組的核酸合成代謝路徑,進(jìn)行了乳清酸磷酸核糖轉(zhuǎn)移酶(OPRT)基因的克隆表達(dá),以及rOPRT的綿羊保護(hù)性效果評(píng)價(jià)。其研究結(jié)果如下:1.對(duì)尤氏泰勒蟲(chóng)、呂氏泰勒蟲(chóng)兩個(gè)種的六個(gè)地方株的蛋白組進(jìn)行核苷酸代謝途徑初步分析,用COG的聚類分析結(jié)果表明,與核苷酸代謝相關(guān)的蛋白有5個(gè),其中4個(gè)參與從頭合成途徑,只有一個(gè)參與補(bǔ)救途徑。其中乳清酸磷酸核糖轉(zhuǎn)移酶、腺苷酸激酶分別為從頭合成途徑、補(bǔ)救途徑的關(guān)鍵酶,具有重要的研究意義。2.克隆和表達(dá)了尤氏泰勒蟲(chóng)oprt基因的功能區(qū)片段,并對(duì)重組蛋白rOPRT的反應(yīng)原性進(jìn)行分析。結(jié)果表明oprt基因的表達(dá)產(chǎn)物為包涵體蛋白,rOPRT除與尤氏泰勒蟲(chóng)陽(yáng)性血清反應(yīng)外,也與呂氏泰勒蟲(chóng)和綿羊泰勒蟲(chóng)陽(yáng)性血清發(fā)生交叉反應(yīng)。證實(shí)oprt同源基因也存在于呂氏泰勒蟲(chóng)和綿羊泰勒蟲(chóng)。3.動(dòng)物保護(hù)性實(shí)驗(yàn)完成了rOPRT對(duì)綿羊免遭尤氏泰勒蟲(chóng)感染的保護(hù)效果評(píng)估。結(jié)果表明,對(duì)照組綿羊發(fā)燒持續(xù)時(shí)間明顯長(zhǎng)于免疫組,與對(duì)照組相比,免疫組綿羊帶蟲(chóng)期明顯縮短。對(duì)照組實(shí)驗(yàn)綿羊分別在攻蟲(chóng)血后第25天、26天、29天出現(xiàn)高蟲(chóng)血癥,而免疫組則分別出現(xiàn)在第26天、32天和33天,明顯比對(duì)照組出現(xiàn)的晚。血紅蛋白含量測(cè)定的結(jié)果顯示,對(duì)照組的血紅蛋白含量(Hb)顯著下降,而實(shí)驗(yàn)組Hb變化較小。同時(shí)紅細(xì)胞計(jì)數(shù)和臨床癥狀觀察,可以看出對(duì)照組貧血嚴(yán)重且持續(xù)時(shí)間長(zhǎng)。4.用ELISA對(duì)2012年5月到2013年6月份期間采自中國(guó)21個(gè)省份、47個(gè)地區(qū)的8至20月齡綿羊和山羊的2351份血清進(jìn)行了檢測(cè),對(duì)我國(guó)羊泰勒蟲(chóng)病的流行情況進(jìn)行調(diào)查,結(jié)果表明全國(guó)各地血清學(xué)陽(yáng)性率存在明顯差異,如甘肅省羊的泰勒蟲(chóng)病血清學(xué)陽(yáng)性率最高,為76%,而河北省血清學(xué)陽(yáng)性率最低,未檢測(cè)到羊泰勒蟲(chóng)感染。
[Abstract]:Sheep Taylor's disease is a disease caused by the parasites of the genus Thaleridae, which is spread by vector ticks, in macrophages, lymphocytes and red blood cells of sheep and goats. It has brought great economic losses to the global sheep industry. In this study, bioinformatics was used to analyze the nucleic acid biosynthesis pathway of sheep Taylor's proteome, and to clone and express the nucleic acid transcriptase (OPRT) gene of whey acid phosphate ribonucleosyltransferase (OPRT). The results of this study are as follows: 1. The nucleotide metabolic pathways of six local strains of Taylor Eurex and Taylor lui were preliminarily analyzed, and the results of COG cluster analysis showed that, There are five proteins related to nucleotide metabolism, four of which are involved in the ab initio pathway, and only one is involved in the rescue pathway, in which whey acid phosphate ribosyltransferase and adenylate kinase are the key enzymes in the ab initio synthesis pathway, respectively. Cloning and expression of the functional region of the oprt gene of Taylor Eurex. The reactivity of the recombinant protein rOPRT was analyzed. The results showed that the expression product of oprt gene was the inclusion body protein rOPRT, which reacted with the positive serum of Taylor's parasite. It was confirmed that oprt homologous gene was also present in Taylor lui and Taylor's sheep. 3. Animal protective experiment was completed to protect sheep from Taylor lucifera infection. The results show that, The duration of fever in the control group was significantly longer than that in the immunized group, and compared with that in the control group, the period of infection in the control group was significantly shorter than that in the control group. The experimental sheep in the control group developed hyperwormemia on the 25th day, the 26th day and the 29th day, respectively. However, the immunized group appeared on the 26th day, 32 days and 33 days, respectively, which was obviously later than that of the control group. The results of hemoglobin content test showed that the hemoglobin content of the control group decreased significantly. The HB of the experimental group was small, and the red blood cell count and clinical symptoms were observed. It can be seen that anemia in the control group is severe and lasting for a long time. ELISA was used to detect 2,351 serum samples of sheep and goats aged 8 to 20 months collected from 21 provinces and 47 regions of China from May 2012 to June 2013. The prevalence of Taylor's disease in sheep in China was investigated. The results showed that there were significant differences in seropositive rate between different parts of the country. For example, the seropositive rate of Taylor's disease in sheep in Gansu Province was the highest (76%), while the seropositive rate in Hebei Province was the lowest. No Taylor's infection was detected in sheep.
【學(xué)位授予單位】：中國(guó)農(nóng)業(yè)科學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S852.7

【共引文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前2條

1 劉慶;鑒別診斷尤氏泰勒蟲(chóng)和呂氏泰勒蟲(chóng)的LAMP方法的建立與初步應(yīng)用[D];東北農(nóng)業(yè)大學(xué);2013年

2 朱丹;我國(guó)部分地區(qū)羊梨形蟲(chóng)病PCR檢測(cè)方法建立及分子流行病學(xué)調(diào)查[D];河南農(nóng)業(yè)大學(xué);2012年

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本文編號(hào)：1557314