INP-2XFlag S
Part name: BBa_K2509020
Short description: INP-2XFlag S
Schematic diagram:
Long description: This part encodes a fusion protein that composed of INP and 2XFlag tag and designed to be expressed at the surface of E.coli. We replaced the 6His tag of INP-6His (BBa_K2509019) with another tag 2XFlag (BBa_K2509007) to build this one. After being transferred with this part, the host E.coli strains is considered to have the ability of binding with anti-Flag magnetic beads.
Source of the part: BBa_K2509019 and BBa_K2509007
Design considerations: we added a stop codon at the end of 2XFlag tag so as to make a C-terminal fusion protein of INP and 2XFlag.
Results:
Dot-Blot result and Western-blot result
This part was expressed correctly in E.coli.
Immunofluorescence (IF) results
Conclusion: The 6His tag was expressed at the surface of E.coli strain as designed.
References:
1. Fan LH, Liu N, Yu MR, Yang ST, Chen HL. Cell surface display of carbonic anhydrase on Escherichia coli using ice nucleation protein for CO₂ sequestration. Biotechnol Bioeng. 2011 Dec; 108(12):2853-64.
2. Eui-Joong Kim and Seung-Ku Yoo. Cell surface display of CD8 ecto domain on Escherichia coli using ice nucleation protein. Biotechnology Techniques. 1998 Mar; 12(3):197–201.
3. Kim EJ, Yoo SK.Cell surface display of hepatitis B virus surface antigen by using Pseudomonas syringae ice nucleation protein. Lett Appl Microbiol. 1999 Nov; 29(5):292-7.
4. Chungjatupornchai W, Fa-aroonsawat S. Translocation of green fluorescent protein to cyanobacterial periplasm using ice nucleation protein. J Microbiol. 2009 Apr; 47(2):187-92.
