Construction Of Venom Gene CDNA Library Of Vespa Velutina Nigrithorax Buysson

Vespa is a vital economic insect. Vespa venom comprises two groups of compounds: proteins and non-proteins. The non-proteins are of little amount while neurotoxins are the main content of proteins and the major acting factors that cause death and paralysis of mammals insects and crustaceans, also have very important clinical applied value at disease therapy of hypersensitivity nerve heart and blood vessel, so it has significant reason to research and exploitation.Master allergen-proteins' genes of vespa venom with big molecular weight were researched by foreign scientists, for example: Phospholipase A1 Hyaluronidase and Antigen 5. We had no new materials about ingredients of Vaspa velutina nigrithorax Buysson venom in details. Habermann suggested that waspas and vespas have no peptides similar to melittin apamin Mast Cell Degranulating and secapin with bees. Zhang Su-fang and Shi Wan-jun referred to venom genes of Apis melifica to synthesized primers, got preproapamin gene prepromelittin gene and precursor of MCD peptide's gene of Vaspa velutina nigrithorax Buysson by RT-PCR in institute of applied entomology of Zhejiang University, and contrasted these genes with Apis melifica's genes, and found there are only a few nucleotides in difference. It is a new route to research ingredients of this venom roundly.Nature venom of vespa is separated from their precursors transferred and machinized by enzymes in different tissues. The relation between complicated transformation and genes is not known. Vespas and bees are different sections, and their venoms have different functions. It is a good subject to research the difference between two venom's ingredients because bee's venom is to defend while vespa's venom hasdefendable and raptorial functions. There are manifold venom peptides in vespa sac, but it is a blank research square about these genes at home.Venom compound of Vaspa velutina nigrithorax Buysson is complicated rare and hard-won. It is essential for research chemical synthesizing and biological synthesizing. If we can get to the bottom of venom's property of physical-chemistry and functionary mechanism, we get a firm foundation to empolder new medicine of disease therapy and pesticide. In order to research venom's biological compound and function of Vaspa velutina nigrithorax Buysson, we used the venom sac as material and constructed the cDNA library of venom sac's genes.Total RNA was extracted from venom sac of Vaspa velutina nigrithorax Buysson and purified by oligo dT magnet separation. The first strand cDNA was synthesized using AMV reverse transcriptase system, and then the second strand by DNA Polymerase I and RNase H. The fragments longer than 50 bp were purified and ligated into pUC118. We analyzed the 780 clones of cDNA library, in which more than 98.5% clones are positive. Most of insert DNA were about 500-900bp. The data showed that cDNA library had reasonably good quality. We analyzed the sequence of 20 clones, and discovered several interesting ORFs. The function of genes must be researched ulteriorly.