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| Proteins, Expression, Isolation and Analysis | The coding sequence of CB (isoform C) obtained from the National Center for Biotechnology Information (NCBI) database (GenBank: X12603.1) was used as a reference. The cloning and expression of the recombinant CB (isoform C) was carried out using a pET-30b(+) vector, which contains and operates using the bacteriophage T7 promoter. The cloning of the coding sequence and the subcloning of the sequence in the pET-30b(+) vector was performed by Genscript USA Inc, using the NotI-HindIII restriction sites. Protein expression was conducted using RosettaTM (DE3)pLysS cells, genotype F- ompT hsdSB(rB- mB- ) gal dcm (DE3) pLysSRARE (CamR), which work as a protease deficient expression host. | Get A Quote |
Crotoxin is a heterodimeric β-neurotoxin isolated from the venom of Crotalus durissus terrificus, consisting of two, non-covalently bound subunits, crotapotin (CA) and crotoxin B (CB). Both subunits present four different isoforms, consequently there are up to 16 different crotoxin complexes, each with different activity levels. The biological activities usually associated with crotoxin include neurotoxicity, myotoxicity and cardiotoxicity, however several other important biochemical and pharmacological effects of crotoxin have been observed, such as the antibacterial, antiviral, antiinflammatory, antitumoral and analgesic activity. In this study we present the production of recombinant crotoxin B (isoform C).... More
