Illegal EcoRI site found at 1813 Illegal PstI site found at 295 Illegal PstI site found at 304 Illegal PstI site found at 1456
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 1813 Illegal PstI site found at 295 Illegal PstI site found at 304 Illegal PstI site found at 1456
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 1813 Illegal BglII site found at 533
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 1813 Illegal PstI site found at 295 Illegal PstI site found at 304 Illegal PstI site found at 1456
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 1813 Illegal PstI site found at 295 Illegal PstI site found at 304 Illegal PstI site found at 1456 Illegal AgeI site found at 1495
1000
COMPATIBLE WITH RFC[1000]
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.
