本文選題：日本血吸蟲(chóng) + 童蟲(chóng)　； 參考：《中南大學(xué)》2010年碩士論文

【摘要】： [目的]從噬菌體十二肽庫(kù)中篩選出與日本血吸蟲(chóng)(Schistosoma japonicum, Sj)童蟲(chóng)表膜特異性結(jié)合的多肽,為研究血吸蟲(chóng)病的治療提供靶向分子。 [方法]利用噬菌體十二肽庫(kù)與Sj活童蟲(chóng)靶分子之間的親和力結(jié)合,經(jīng)過(guò)三輪吸附-洗脫-擴(kuò)增的篩選過(guò)程,從末次回收的結(jié)合噬菌體中隨機(jī)挑取20個(gè)陽(yáng)性克隆,首先對(duì)其進(jìn)行測(cè)序；隨后利用免疫組化、Western blot分析等方法觀察目標(biāo)噬菌體在體外與童蟲(chóng)表膜的結(jié)合能力,并通過(guò)動(dòng)物實(shí)驗(yàn)將目標(biāo)噬菌體回輸?shù)揭迅腥狙x(chóng)的動(dòng)物體內(nèi),分別回收肝臟和血吸蟲(chóng)童蟲(chóng),分別洗脫與肝臟和童蟲(chóng)結(jié)合的噬菌體,進(jìn)行統(tǒng)計(jì)學(xué)分析,驗(yàn)證其靶向性。 [結(jié)果]經(jīng)3輪篩選后,噬菌體回收率從第1輪的0.77×10-8增加到第3輪的0.75×10-5,說(shuō)明噬菌體得到了有效富集。DNA測(cè)序后發(fā)現(xiàn),20個(gè)陽(yáng)性噬菌體克隆中的15個(gè)克隆的插入序列均為QSFASLTNPRVL,將這些噬菌體克隆命名為ZW-15。免疫組化、Western blot均顯示ZW-15噬菌體的該插入序列能與Sj童蟲(chóng)表膜有效結(jié)合。體內(nèi)回輸試驗(yàn)比較ZW-15及M13KE空載噬菌體的回收量,兩者從單位重量蟲(chóng)體中的回收量差異顯著(P10.01),而兩者從單位重量肝臟中的回收量差別無(wú)統(tǒng)計(jì)學(xué)意義(P20.05),證實(shí)該ZW-15噬菌體的插入序列短肽能有效地靶向結(jié)合于體內(nèi)Sj童蟲(chóng)表膜。 [結(jié)論]本研究篩選獲得陽(yáng)性噬菌體克隆ZW-15所展示的短肽QSFASLTNPRVL能有效地靶向結(jié)合Sj童蟲(chóng)表膜,將為進(jìn)一步研究血吸蟲(chóng)病的治療提供靶向分子。
[Abstract]:[objective] to screen the peptides specifically binding to Schistosoma japonicum (SjjS) surface membrane from phage dodeceptide library, and to provide targeted molecules for the treatment of schistosomiasis. [methods] by using the affinity binding between phage dodeceptide library and Sj live child worm target molecule, 20 positive clones were randomly selected from the last recovered phage after three rounds of screening process of adsorption, elution and amplification. Then the binding ability of the target phage to the surface membrane of the child worm in vitro was observed by immunohistochemistry and Western blot analysis, and the target phage was transferred back to the infected animal by animal experiment. The liver and Schistosoma japonicum were recovered, the phage which was bound to the liver and the child worm was eluted, and the targeting of the bacteriophage was verified by statistical analysis. [results] after three rounds of screening, The phage recovery increased from 0.77 脳 10 ~ (-8) in the first round to 0.75 脳 10 ~ (-5) in the third round. The results showed that 15 of the 20 positive phage clones were inserted into QSFASLTNPRVL. the phage clones were named ZW-15. Immunohistochemical analysis showed that the insertion sequence of ZW-15 phage could effectively bind to the surface membrane of Sj child worm. The recoveries of ZW-15 and M13KE were compared in vivo. There was a significant difference in the recovery amount between the two species (P 10.01), but there was no significant difference in the recovery amount between the two species from the liver (P 20.05). It was confirmed that the inserted short peptide of the ZW-15 phage could be effectively targeted to the surface membrane of Sj children's worm in vivo. [conclusion] in this study, the short peptide QSFASLTNPRVL displayed by the positive phage clone ZW-15 can be effectively targeted to the surface membrane of Sj children's worm, which will provide a target molecule for further study on the treatment of schistosomiasis.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R392

【參考文獻(xiàn)】

中國(guó)期刊全文數(shù)據(jù)庫(kù) 前10條

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2 歐陽(yáng)理,王慶林,曾憲芳,周金春,易新元;從隨機(jī)多肽庫(kù)中篩選日本血吸蟲(chóng)抗原模擬表位誘導(dǎo)保護(hù)性免疫的研究[J];中國(guó)熱帶醫(yī)學(xué);2002年02期

3 金怡,錢e，

本文編號(hào)：1933992