本文選題：羊泰勒蟲　切入點：蛋白組OPRT　出處：《中國農(nóng)業(yè)科學院》2015年碩士論文　論文類型：學位論文

【摘要】：羊泰勒蟲病是由媒介蜱傳播的泰勒科泰勒屬的原蟲寄生于綿羊和山羊的巨噬細胞、淋巴細胞和紅細胞內(nèi)而引起的疾病的總稱,它給全球養(yǎng)羊業(yè)帶來了巨大的經(jīng)濟損失。本研究運用生物信息學初步分析了羊泰勒蟲蛋白組的核酸合成代謝路徑,進行了乳清酸磷酸核糖轉(zhuǎn)移酶(OPRT)基因的克隆表達,以及rOPRT的綿羊保護性效果評價。其研究結(jié)果如下:1.對尤氏泰勒蟲、呂氏泰勒蟲兩個種的六個地方株的蛋白組進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,其中4個參與從頭合成途徑,只有一個參與補救途徑。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、補救途徑的關(guān)鍵酶,具有重要的研究意義。2.克隆和表達了尤氏泰勒蟲oprt基因的功能區(qū)片段,并對重組蛋白rOPRT的反應(yīng)原性進行分析。結(jié)果表明oprt基因的表達產(chǎn)物為包涵體蛋白,rOPRT除與尤氏泰勒蟲陽性血清反應(yīng)外,也與呂氏泰勒蟲和綿羊泰勒蟲陽性血清發(fā)生交叉反應(yīng)。證實oprt同源基因也存在于呂氏泰勒蟲和綿羊泰勒蟲。3.動物保護性實驗完成了rOPRT對綿羊免遭尤氏泰勒蟲感染的保護效果評估。結(jié)果表明,對照組綿羊發(fā)燒持續(xù)時間明顯長于免疫組,與對照組相比,免疫組綿羊帶蟲期明顯縮短。對照組實驗綿羊分別在攻蟲血后第25天、26天、29天出現(xiàn)高蟲血癥,而免疫組則分別出現(xiàn)在第26天、32天和33天,明顯比對照組出現(xiàn)的晚。血紅蛋白含量測定的結(jié)果顯示,對照組的血紅蛋白含量(Hb)顯著下降,而實驗組Hb變化較小。同時紅細胞計數(shù)和臨床癥狀觀察,可以看出對照組貧血嚴重且持續(xù)時間長。4.用ELISA對2012年5月到2013年6月份期間采自中國21個省份、47個地區(qū)的8至20月齡綿羊和山羊的2351份血清進行了檢測,對我國羊泰勒蟲病的流行情況進行調(diào)查,結(jié)果表明全國各地血清學陽性率存在明顯差異,如甘肅省羊的泰勒蟲病血清學陽性率最高,為76%,而河北省血清學陽性率最低,未檢測到羊泰勒蟲感染。
[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.
【學位授予單位】：中國農(nóng)業(yè)科學院
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S852.7

【共引文獻】

相關(guān)碩士學位論文 前2條

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

2 朱丹;我國部分地區(qū)羊梨形蟲病PCR檢測方法建立及分子流行病學調(diào)查[D];河南農(nóng)業(yè)大學;2012年

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本文編號：1557314