Part:BBa_K4815010
Description
E. coli heat-labile enterotoxin subunit B (LTB) is an important oral vaccine adjuvant widely used to prevent various diseases such as cholera, traveler’s diarrhea, and E. coli infection. Compared traditional LTB production methods, such as the E. coli expression system, LTB produced by brewing yeast is purer and does not contain residual host cell materials and endotoxins. It has good safety and immunogenicity. At the same time, LTB produced by brewing yeast shows good immunogenicity in oral vaccines, effectively activating the immune system and inducing specific immune responses. However, limited expression rate of LTB in yeasts have remaining a grate problem for a long time and the promoter is an important element in gene expression regulation. Combined with our pymaker, we predicted and provided a series of suitable efficient promoters, and we successfully applied the predicted promoters to express LTB in brewing yeast by combining the predicted results with the gene expression system of brewing yeast. To detect the expression rate of the LTB, we fused LTB to the green fluorescent protein. On the one hand, FACS analysis can be easily applied to measure cell fluorescence to detect the expression rate of the LTB, on the other hand, the LTB-eGFP fusion protein are more accessible to run western blot choosing the antibody of GFP.
Origin
Heat-labile enterotoxin subunit B (LTB) which is expressed by E. coli is one of the most popular oral vaccine adjuvants and intestine adsorption enhancers. We fused LTB to eGFP and we tested our expression system for eGFP as a model reporter protein.
Composite
LTB-eGFP consists two parts: the Heat-labile enterotoxin subunit B and the green fluorescent protein. We follow the procedure in the related research [1],combine LTB(the core sequence of BBa_K1074005) and eGFP(the core sequence of BBa_E0040) with 4 amino acids 'CSCS' as a fusion protein. The Heat-labile enterotoxin subunit B is the oral vaccine adjuvants and intestine adsorption enhancers. The green fluorescent protein act as the reporter protein.
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