Part:BBa_K5382120:Design
InaK-linker-Im7_Cell membrane anchoring protein and immune protein complex
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1800
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 722
Illegal NgoMIV site found at 962
Illegal NgoMIV site found at 1130
Illegal AgeI site found at 517 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 692
Illegal BsaI.rc site found at 1182
Illegal SapI.rc site found at 326
Design considerations
Experimental result
We transferred pET23a-linker-Im7 into Escherichia coli BL21 and purified it to meet the use requirements. The results were as follows:
Figure 1. EcN engineered bacteria induced expression of InaK-Im7 fusion protein. 1, 3, 5 are before induction, 2, 4, 6 are after induction with IPTG.
It can be seen that the purified protein concentration and purity are high, and subsequent incubation binding and fluorescence confocal experiments can be conducted to verify the binding of Inak-Im7 and CL7-sfGFP (see the engineer and result section of wiki for the results).
Source
The InaK is an ice nucleated protein from Pseudomonas syringae KCTC1832.
The linker is composed of Gly and Ser.
The Im7 is an immune protein derived from E. coli.
The source of the composite parts is artificially constructed plasmids(pCold-Inak-Im7).