广西扇棘单睾吸虫5.8S rRNA序列和二级结构研究

发布时间：2019-01-08 20:46

【摘要】： 目的:研究广西扇棘单睾吸虫和其它人体吸虫5.8S rRNA序列和二级结构,探讨人体吸虫二级结构的特征,揭示单睾吸虫之间的亲缘关系。 方法:采集广西扶绥县扇棘单睾吸虫,提取其DNA,PCR扩增5.8S rDNA序列并测序。另外选用Genbank登录的7种人体吸虫5.8S rDNA序列进行研究,包括:扇棘单睾吸虫、钩棘单睾吸虫、巨片形吸虫、裂体吸虫、獾似颈吸虫、卷棘口吸虫和猫后睾吸虫。应用DNAman软件把上述人体吸虫5.8SrDNA基因转换成5.8S rRNA。应用RNAstructure3.2软件,根据最小自由能原理,采用Zuker算法,构建5.8S rRNA分子二级结构。应用SPSSv14.0软件对5.8S rRNA一级结构和二级结构组成进行统计分析,人工比对分析部分人体吸虫5.8S rRNA序列和二级结构特征。 结果:对广西扇棘单睾吸虫与Dzikowski等报道的扇棘单睾吸虫5.8SrRNA序列进行比较,显示第81、83、84、129、143位碱基发生变异,但这些突变碱基均位于5.8S rRNA二级结构未配对碱基区,碱基突变并未影响5.8S rRNA二级结构。对广西扇棘单睾吸虫与钩棘单睾吸虫5.8S rRNA序列进行比较,发现既存在未配对碱基区的变异又存在配对碱基区的变异,配对区碱基的变异使两者5.8S rRNA二级结构呈现较大差异。对复殖目部分吸虫5.8S rRNA二级结构进行比对分析,结果显示:5.8S rRNA二级结构具有相同的基本结构模式;Ⅰ分支具有1个保守模序为5′-GUGUCGAUG:CAUCGACAU-3′(钩棘单睾吸虫为5′-GUGUCGAUG:CAUCGAUAU-3′)。Ⅱ分支具有1个保守发卡结构,该发卡结构的茎是由9个碱基对组成的保守模序,S.sp保守模序为5′-GGCUACGGG:CCUGUGGCC-3′,其它7种保守模序为5′-GGCCAUGGG:CCUGUGGCC-3′;发卡环为富含尿嘧啶的6元环。在8种人体吸虫中,除裂体吸虫和猫后睾吸虫外,其它6种吸虫5.8SrRNA 5′端均有1段单链序列。在8种人体吸虫中,除钩棘单睾吸虫外,其余吸虫Ⅰ分支均有保守模序5′-GCAG:CUGC-3′。钩棘单睾吸虫和裂体吸虫与其它6种吸虫具有不同的H部和多分支环。巨片形吸虫、獾似颈吸虫和卷棘口吸虫Ⅰ分支尾端具有一个保守发卡结构。碱基的替代、丢失和插入多发生于5.8S rRNA二级结构单链区,双链区的替代、丢失和插入则往往会引起二级结构的较大变异。 结论:本研究首次获得广西扇棘单睾吸虫5.8S rRNA基因序列,发现人体吸虫5.8S rRNA二级结构的一些特征。根据单睾吸虫二级结构特征,本研究还发现广西扇棘单睾吸虫与Dzikowski等报道的扇棘单睾吸虫亲缘关系比较近,与钩棘单睾吸虫亲缘关系比较远。
[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，

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