Similar protocols

Pipeline publication

[…] ed for 2 days by electrical stimulation. Total RNA was prepared from the glands by using TRIzol reagent (Invitrogen). Poly (A) mRNA was purified by using a Poly (A) Tract mRNA isolation system (Promega). The cDNA library was constructed with the Superscript plasmid system cDNA library construction kit (Gibco/BRL). cDNAs were cloned into the pSPORT1 plasmid (Gibco/BRL) and transformed into Escherichia coli DH5α cells. cDNA clones were randomly chosen and sequenced to obtain a reliable representation of the toxin content in the venom gland. Positive clones were identified by using an ABI Prism 377XL DNA sequencer with a universal T7 promoter primer., Sequence analysis was carried out by using BLASTX (http://blast.ncbi.nlm.nih.gov/Blast.cgi), DNAMAN, and GENRUNR. The secondary structure was analyzed by using the online program Heliquest (http://heliquest.ipmc.cnrs.fr/cgi-bin/ComputParams.py) and measured by circular dichroism (CD) spectroscopy. The CD assay was performed at room temperature at the UV range of 190–250 nm at a concentration of 0.1 mg/mL in: (i) water, (ii) 30% TFE/H2O, or (iii) 70% TFE/H2O. Spectra were collected from three separate recordings., The peptides used in this study were synthesized by GL Biochem (Shanghai, China) with amidated C-terminus, and with a purity of >95%. Staphylococcus aureus AB94004, S. aureus ATCC25923, S. aureus ATCC6538, S. aureus AB208193, S. epidermidis AB208187, S. epidermidis AB208188, Micrococcus luteus AB93113, Bacillus subtilis AB91021, E. coli AB94012, E. coli ATCC25922, Pseudomo […]

Pipeline specifications

Software tools BLASTX, DNAMAN, HELIQUEST