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===Design Notes=== | ===Design Notes=== | ||
Revision as of 17:54, 8 October 2023
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Design Notes
In our design, we aimed to introduce the gene fragment encoding an active Cry3A-like toxin into Escherichia coli using the pET30a vector as a carrier, in order to confer upon it the ability to produce Cry3A-like toxin. To achieve secretion expression, we added a signal peptide sequence, OmpA, to the N-terminus of Cry3A-like toxin. This signal peptide was included to guide the transport of Cry3A-like toxin to the extracellular space. OmpA is a commonly used signal peptide in Escherichia coli that has been verified for the secretion expression of foreign proteins. Additionally, we fused a 6×His tag to the C-terminus of Cry3A-like toxin to facilitate subsequent protein purification and enable specific characterization experiments using Western blot.
Source
We obtained the sequence of the active Cry protoxin of UTD-001 from the original literature. After codon optimization, we entrusted GUANGZHOU IGE BIOTECHNOLOGY LTD to synthesize the gene fragment OmpA-active Cry protoxin of UTD-001.
References
[1] Lee A. Bulla, Jr.Mehmet Candas Formicidae (ant) control using Bacillus thuringiensis toxin US 6,551,800B1[P]. 2003-04-22.