Abstract: 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 43 157 unigene genes were obtained by sequencing, 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 predation 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.