Vespa crabro is a cosmopolitan social wasp species whereas Vespa analis is commonly found in Asia. Both species are widely distributed in Korea and known to be aggressive when disturbed, resulting in frequent sting accidents. Although major venom components of well known Vespa wasps have been reported, no comparative transcriptomic analysis of venom gland between V. crabro and V. analis has been conducted to date. To investigate the differences in venom properties between these two wasps, total RNA was extracted from each venom gland and used for RNA-sequencing. A total of 31 venom-specific genes were identified in both venom gland transcriptomes but their expression profiles were different between V. crabro and V. analis. Venom allergen 5, premastoparan A and phospholipase A were the top three genes that were most prevalently transcribed in the venom gland of V. crabro, and their transcription rates were 902-, 112- and 4164-fold higher compared with V. analis, respectively, as judged by FPKM values. Their differential transcription profiles were confirmed by quantitative real-time PCR. In the venom gland of V. analis, however, premastoparan A was most abundantly transcribed gene, followed by calponin and tropomysin. In general, most venom-specific genes were more abundantly expressed in V. crabro but some genes exhibited higher transcription rates in V. analis, including muscle LIM protein, troponin, paramyosin, calponin, etc. Our findings suggest that V. crabro produce venom with much more enriched venom components, thereby with higher toxicity compared with V. analis.