【摘要】： 目的:研究廣西扇棘單睪吸蟲和其它人體吸蟲5.8S rRNA序列和二級(jí)結(jié)構(gòu),探討人體吸蟲二級(jí)結(jié)構(gòu)的特征,揭示單睪吸蟲之間的親緣關(guān)系。 方法:采集廣西扶綏縣扇棘單睪吸蟲,提取其DNA,PCR擴(kuò)增5.8S rDNA序列并測(cè)序。另外選用Genbank登錄的7種人體吸蟲5.8S rDNA序列進(jìn)行研究,包括:扇棘單睪吸蟲、鉤棘單睪吸蟲、巨片形吸蟲、裂體吸蟲、獾似頸吸蟲、卷棘口吸蟲和貓后睪吸蟲。應(yīng)用DNAman軟件把上述人體吸蟲5.8SrDNA基因轉(zhuǎn)換成5.8S rRNA。應(yīng)用RNAstructure3.2軟件,根據(jù)最小自由能原理,采用Zuker算法,構(gòu)建5.8S rRNA分子二級(jí)結(jié)構(gòu)。應(yīng)用SPSSv14.0軟件對(duì)5.8S rRNA一級(jí)結(jié)構(gòu)和二級(jí)結(jié)構(gòu)組成進(jìn)行統(tǒng)計(jì)分析,人工比對(duì)分析部分人體吸蟲5.8S rRNA序列和二級(jí)結(jié)構(gòu)特征。 結(jié)果:對(duì)廣西扇棘單睪吸蟲與Dzikowski等報(bào)道的扇棘單睪吸蟲5.8SrRNA序列進(jìn)行比較,顯示第81、83、84、129、143位堿基發(fā)生變異,但這些突變堿基均位于5.8S rRNA二級(jí)結(jié)構(gòu)未配對(duì)堿基區(qū),堿基突變并未影響5.8S rRNA二級(jí)結(jié)構(gòu)。對(duì)廣西扇棘單睪吸蟲與鉤棘單睪吸蟲5.8S rRNA序列進(jìn)行比較,發(fā)現(xiàn)既存在未配對(duì)堿基區(qū)的變異又存在配對(duì)堿基區(qū)的變異,配對(duì)區(qū)堿基的變異使兩者5.8S rRNA二級(jí)結(jié)構(gòu)呈現(xiàn)較大差異。對(duì)復(fù)殖目部分吸蟲5.8S rRNA二級(jí)結(jié)構(gòu)進(jìn)行比對(duì)分析,結(jié)果顯示:5.8S rRNA二級(jí)結(jié)構(gòu)具有相同的基本結(jié)構(gòu)模式;Ⅰ分支具有1個(gè)保守模序?yàn)?′-GUGUCGAUG:CAUCGACAU-3′(鉤棘單睪吸蟲為5′-GUGUCGAUG:CAUCGAUAU-3′)。Ⅱ分支具有1個(gè)保守發(fā)卡結(jié)構(gòu),該發(fā)卡結(jié)構(gòu)的莖是由9個(gè)堿基對(duì)組成的保守模序,S.sp保守模序?yàn)?′-GGCUACGGG:CCUGUGGCC-3′,其它7種保守模序?yàn)?′-GGCCAUGGG:CCUGUGGCC-3′;發(fā)卡環(huán)為富含尿嘧啶的6元環(huán)。在8種人體吸蟲中,除裂體吸蟲和貓后睪吸蟲外,其它6種吸蟲5.8SrRNA 5′端均有1段單鏈序列。在8種人體吸蟲中,除鉤棘單睪吸蟲外,其余吸蟲Ⅰ分支均有保守模序5′-GCAG:CUGC-3′。鉤棘單睪吸蟲和裂體吸蟲與其它6種吸蟲具有不同的H部和多分支環(huán)。巨片形吸蟲、獾似頸吸蟲和卷棘口吸蟲Ⅰ分支尾端具有一個(gè)保守發(fā)卡結(jié)構(gòu)。堿基的替代、丟失和插入多發(fā)生于5.8S rRNA二級(jí)結(jié)構(gòu)單鏈區(qū),雙鏈區(qū)的替代、丟失和插入則往往會(huì)引起二級(jí)結(jié)構(gòu)的較大變異。 結(jié)論:本研究首次獲得廣西扇棘單睪吸蟲5.8S rRNA基因序列,發(fā)現(xiàn)人體吸蟲5.8S rRNA二級(jí)結(jié)構(gòu)的一些特征。根據(jù)單睪吸蟲二級(jí)結(jié)構(gòu)特征,本研究還發(fā)現(xiàn)廣西扇棘單睪吸蟲與Dzikowski等報(bào)道的扇棘單睪吸蟲親緣關(guān)系比較近,與鉤棘單睪吸蟲親緣關(guān)系比較遠(yuǎn)。
[Abstract]:Objective: to study the 5.8s rRNA sequence and secondary structure of Clonorchis fannis and other human paragonimiasis in Guangxi, and to explore the characteristics of secondary structure of human trematorchiasis, and to reveal the relationship between Clonorchis sinensis. Methods: the 5.8s rDNA sequence was amplified by DNA,PCR from Clonorchis fan in Fusui County, Guangxi, and sequenced. In addition, the 5.8s rDNA sequence of 7 species of human trematodes recorded by Genbank was used to study them, including: Monoclonorchis fan, clonorchiasis hook, clonorchiasis monocystis, clonorchiasis, clonorchiasis sinensis, paragonimiasis sinensis, paragonimus capillaris and clonorchiasis posterior to cat. Transformation of the 5.8SrDNA gene from the human trematorrhea to 5.8s rRNA. by DNAman software Based on the principle of minimum free energy and Zuker algorithm, the secondary structure of 5.8S rRNA molecule was constructed by using RNAstructure3.2 software. The primary and secondary structures of 5.8S rRNA were statistically analyzed by SPSSv14.0 software. The 5.8S rRNA sequences and secondary structure characteristics of some human trematodes were analyzed by artificial comparison. Results: by comparing the 5.8SrRNA sequences of Clonorchis fannicans and Dzikowski, the results showed that the 81h83p8129143 bases were mutated, but they were located in the unpaired region of 5.8S rRNA secondary structure. Base mutation did not affect the secondary structure of 5.8S rRNA. By comparing the 5.8s rRNA sequences of Clonorchis fannis and Clonorchis hook Monochidism in Guangxi, it was found that there were variations in both unpaired and paired base regions. The secondary structure of 5.8S rRNA showed great difference due to the variation of the base in the pairing region. The secondary structure of 5.8S rRNA was compared and analyzed. The results showed that the secondary structure of 5.8S rRNA had the same basic structure pattern. Branch 鈪，

本文編號(hào)：2405098