Proteomic Analysis Of The Venom Collected From Latrodectus Tredecimguttatus By Electrostimulation

Part I : Hibernation and early indoor-rearing of the baby spiders of Latrodectus tredecimguttatusLatrodectus tredecimguttatus, "black widow spider" or "black venomous spider" , is one of medium-sized venomus spiders, which belong to Phylum Arthopoda, Arachnidea, Araneae, Theridiidae, genus Latrodectus. The source of toxins limited the research on it due to the very low venom yield of the spider. Therefore, indoor-rearing of Latrodectus tredecimguttatus is an effective method for resolving the source of toxins and protecting the spider resource. Hibernation and early rearing of baby spiders of Latrodectus tredecimguttatus is crutical to indoor-rearing of the spider. In this article, The hibernation conditions and early growth and development of baby spiders of L. tredecimguttatus were investigated in detail. The results indicate that the optimal temperature for hibernation of baby spider is around 4°C. The optimal length of hibernation is 6-7 months. And the egg bags must be placed at room temperature a period of time prior to being preserved at low temperature. After these baby spiders were reared for a month with Tenebtio molitor L. in 50%~55% of relative humidity and 25°C~30°C, the average body weights of baby spider increased by more than 40-fold, with survival ratio greater than 80%. PartⅡ: Proteomic analysis of venom collected from Latrodectus tredecimguttatus by electrostimulationBlack widow spider is one of the most poisonous spiders in the world. Up to now, there was not any report on its venom proteome. In this paper, we employed combinative proteomic strategies to analyze the veom collected from living spider Latrodectus tredecimgtuttatus by electrostimulation. The combinative proteomic strategies including the following two different approaches: (ⅰ) one-dimensional SDS/PAGE plus ESI-Q-TOF, (ⅱ) shotgun-fractionation plus ESI-Q-TOF. The experiments demonstrated that the venom has a high aboundance of high-molecular-weight proteins (around 100kDa), and the content of proteins and peptides below 10 kDa is low. Totals of 152 venomous proteins were identified using cross-species searching. Besides typtical black widow spider toxins (latrotoxins), several hydrolases and other especial proteins were found in the venom, which were speculated to be related to the latrodectism. In addition, 41 venomous proteins identified were of unknown function, and on average about half of high-quality MS/MS spectra gave no signification search results, most likely due to the lack of a specific database for spider venom.