7种榕小蜂的Argonaute蛋白基因的分子进化
发布时间：2018-07-18 12:10
【摘要】：Argonaute蛋白是一类与小RNA相互作用而起调控作用的蛋白。Argonaute蛋白在很多生物体包括植物和动物中都有发现,而且在所有生物体中都极其保守。Argonaute蛋白与小RNA结合,起基因调控的作用。它可以行核酸内切酶的作用和RNA干扰作用,参与表观遗传调控、转录和翻译的调控,因此还对胚胎发育过程有着重要作用。Argonaute蛋白在榕小蜂(膜翅目:小蜂总科)暂无相关研究报道。因此不同榕小蜂物种之间Argonaute蛋白基因的进化关系,以及所有Argonaute蛋白基因经受的选择压力等问题是本研究所感兴趣的。本论文首先通过设计简并引物来扩增Argonaute蛋白的保守片段,再基于保守片段设计hiTAIL-PCR的引物来得到长片段的Argonaute蛋白基因的序列。对于有基因组序列和(或)转录组数据的物种,如对叶榕传粉榕小蜂Ceratosolen solmsi和非传粉榕小蜂Walkerella sp.,还通过手工基因注释和组装并辅以实验验证获得所有Argonaute蛋白基因的序列。结果一共获得了来自七种榕小蜂的16条Argonaute蛋白基因的长片段序列。针对所有已获得的长片段序列,用软件MEGA6.0构建基因树,然后用软件PAML的codeml程序中的位点模型对榕小蜂的Argonaute蛋白基因所受的选择压力进行分析。结果显示:1,除对叶榕传粉榕小蜂C.solmsi缺失PIWI蛋白亚家族的基因而只有AGO蛋白亚家族的Ago2、Ago2-like之外,其它榕小蜂物种都有2-4个Argonaute蛋白基因,既有AGO蛋白亚家族成员,也有PIWI蛋白亚家族成员;2,选择压力分析中,位点模型中的3对比较模型,M0/M3、M1/M2、M7/M8,都没有检测到正选择位点,而且各个分支都受到净化作用。结合各基因所编码蛋白的保守结构域的预测结果,佐证了Argonaute蛋白的保守性。但是C.solmsi缺失PIWI蛋白亚家族的编码基因也因此实在是一个暂时难以解释的问题。
[Abstract]:Argonaute protein is a class of proteins that interact with small RNAs and play the role of gene regulation in many organisms, including plants and animals, and it is extremely conserved in all organisms. It can perform endonuclease and RNA interference, participate in epigenetic regulation, transcription and translation regulation, Therefore, there are no related studies on the role of Argonaute protein in the development of Ficus figs (Hymenoptera: Cercidae). Therefore, the evolutionary relationship of Argonaute protein genes among different species of figs and the selection pressure of all Argonaute genes are of interest to this study. In this paper, degenerate primers were designed to amplify the conserved fragments of Argonaute protein, and then the primers of hiTAIL-PCR were designed to obtain the sequences of long fragments of Argonaute protein gene. For species with genomic sequences and / or transcriptome data, such as Ceratosolen solmsi and Walkerella sp. of fig pollinators, all Argonaute gene sequences were obtained by manual gene annotation and assembly and assisted with experimental verification. Results A total of 16 Argonaute gene fragments were obtained from seven fig wasps. For all the obtained long fragment sequences, the gene tree was constructed by using the software MEGA6.0, and then the selection pressure of the Argonaute protein gene of the fig wasp was analyzed by using the locus model in the codeml program of the software PAML. The results showed that except for the deletion of the PIWI protein subfamily gene to C. solmsi of the fig pollinated fig wasp and only the Ago2Ago2-like gene of the ago protein subfamily, all the other figs had 2-4 Argonaute protein genes, including the ago protein subfamily members. There are also PIWI protein subfamily members. In the selective pressure analysis, three pairs of comparison models of M0 / M3 / M1 / M2M7 / M8 have not detected positive selection sites, and each branch has been purified. The predicted results of conserved domain of the proteins encoded by each gene confirmed the conserved nature of Argonaute protein. However, the deletion of PIWI subfamily genes by C. solmsi is a difficult problem for the time being.
【学位授予单位】：河北大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：Q963
本文编号：2131860
[Abstract]:Argonaute protein is a class of proteins that interact with small RNAs and play the role of gene regulation in many organisms, including plants and animals, and it is extremely conserved in all organisms. It can perform endonuclease and RNA interference, participate in epigenetic regulation, transcription and translation regulation, Therefore, there are no related studies on the role of Argonaute protein in the development of Ficus figs (Hymenoptera: Cercidae). Therefore, the evolutionary relationship of Argonaute protein genes among different species of figs and the selection pressure of all Argonaute genes are of interest to this study. In this paper, degenerate primers were designed to amplify the conserved fragments of Argonaute protein, and then the primers of hiTAIL-PCR were designed to obtain the sequences of long fragments of Argonaute protein gene. For species with genomic sequences and / or transcriptome data, such as Ceratosolen solmsi and Walkerella sp. of fig pollinators, all Argonaute gene sequences were obtained by manual gene annotation and assembly and assisted with experimental verification. Results A total of 16 Argonaute gene fragments were obtained from seven fig wasps. For all the obtained long fragment sequences, the gene tree was constructed by using the software MEGA6.0, and then the selection pressure of the Argonaute protein gene of the fig wasp was analyzed by using the locus model in the codeml program of the software PAML. The results showed that except for the deletion of the PIWI protein subfamily gene to C. solmsi of the fig pollinated fig wasp and only the Ago2Ago2-like gene of the ago protein subfamily, all the other figs had 2-4 Argonaute protein genes, including the ago protein subfamily members. There are also PIWI protein subfamily members. In the selective pressure analysis, three pairs of comparison models of M0 / M3 / M1 / M2M7 / M8 have not detected positive selection sites, and each branch has been purified. The predicted results of conserved domain of the proteins encoded by each gene confirmed the conserved nature of Argonaute protein. However, the deletion of PIWI subfamily genes by C. solmsi is a difficult problem for the time being.
【学位授予单位】：河北大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：Q963
【参考文献】
相关期刊论文 前2条
1 Hsueh-Yen Ku;Haifan Lin;;PIWI proteins and their interactors in piRNA biogenesis,germline development and gene expression[J];National Science Review;2014年02期
2 罗丽娟,施季森;一种DNA侧翼序列分离技术——TAIL-PCR[J];南京林业大学学报(自然科学版);2003年04期
，本文编号：2131860
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