Despite the evolutional distance between wasp and amphibian, vespid chemotactic peptide (VCP), an important component of wasp venom, are found sharing remarkable similarities with the temporin antimicrobial peptides (AMPs) from Ranid frog, Amolops loloensis. Not only their amino acid sequences are highly similar, but they are both microbe-killing and can induce the cellular chemotactic response. However, whether the two peptides possess identical biosynthesis pathway was still not clear due to the unsolved gene sequence of VCP putative precursor. In this paper, a cDNA encoding one of VCP precursors was cloned from the venom sac cDNA library of the wasp, Vespa magnifica (Smith), and the corresponding native VCP was purified from the venoms. It was shown that the VCP precursor highly resembled temporin precursor not only in the sequence size but also in the sequences of their corresponding mature peptides. However, the enzyme-cutting sites and the possible processing enzymes for both peptides were different, which for VCP were dipeptidyl peptidase IV and trypsin-like proteases, while for temporin were only trypsin-like protease. The current results suggested that the biosynthesis mode of VCP was different from that of temporin AMP, even though the two mature peptides were similar in many ways. It is also the first report about VCP precursor from wasp venom.