缘漏斗蛛(Agelena limbata)毒腺转录组学研究

Transcriptomic Research of the Venom Gland of Agelena limbata

为了探究蜘蛛毒素多样性,获取毒素基因序列,进一步研究蜘蛛毒液的成分与应用,本研究在对缘漏斗蛛(Agelena limbata)毒腺进行解剖后采用高通量测序技术对缘漏斗蛛毒腺进行转录组测序,并结合生物信息学方法对转录组进行分析。测序共获得43157个单基因(unigene),经NR(non⁃redundant protein sequence)数据库注释比对,与食母穹蛛(Stegodyphus du⁃micola)同源性最好。GO(the gene ontology resource)富集中蛋白复合物和免疫系统进程所占比例最多。KEGG(Kyoto encyclo⁃pedia of genes and genomes)数据库中与代谢相关的通路占比最高。Uniport数据库中比对的结果显示,毒腺中蛋白质的主要作用为抗菌、杀虫、阻断神经通路、阻断钠离子通道等。简单重复序列(simple sequence repeat,SSR)位点中T/A、AT/TA与TG/CA出现的频率最高。通过单基因的功能注释,初步了解蜘蛛在捕食时毒腺的基因表达情况、基因主要行使的功能及参与的代谢途径,为了解蜘蛛捕食过程提供了基础。此外,根据序列同源性推断结果,发现缘漏斗蛛毒腺毒素中富含半胱氨酸毒素多肽及酶类,还有一些在生物医药领域也发挥重要作用的酶类,例如金属蛋白酶等。综上所述,缘漏斗蛛毒腺中含有丰富的毒素基因,本研究为后续的研究提供了转录组数据来源。

In order to explore the diversity of toxin information of spiders and obtain toxin⁃related gene sequences for the further study and application of spider venom,high throughput sequencing technology was used to sequence the transcriptome of the venom glands of Agelena limbata,and bioinformatic method was used to analyze the transcriptome.A total of 43157 unigene genes were obtained by se⁃quencing,and the best homology was found with Stegodyphus dumicola by NR(Non⁃Redundant Protein Sequence)Database annotation.The proportion of protein complexes and immune system processes is the highest during GO⁃enrichment.The pathways related to meta⁃bolism accounted for the highest proportion in KEGG(Kyoto Encyclopedia of Genes and Genomes)database.The results of comparison in Uniport database show that the main function of proteins in venom glands were antibacterial,insecticidal,blocking nerve pathways,blocking sodium ion channels,etc.T/A,AT/TA and TG/CA appeared most frequently in SSR(simple sequence repeat)loci.Through the functional annotation of unigene,we have preliminarily understood the gene expression of the venom glands of spiders during predation,the main functions of genes and the metabolic pathways involved,which provide a basis for us to understand the pre⁃dation process of spiders.In addition,according to the sequence homology,it was found that the venom glands of Agelena limbata is rich in cysteine toxin polypeptides and enzymes,as well as some enzymes that also play an important role in the field of biomedicine,such as metalloproteinases.In conclusion,the venom glands of Agelena limbata contain abundant toxin genes,which provide a source of transcriptome data for subsequent studies.