Difference between revisions of "Part:BBa K4614002"

Revision as of 06:44, 6 October 2023

Lnak-R5

We used T7 promoter to induce IPN and R5(part BBa_K461400) expression, IPN, a carrier protein from the genome of Pseudomonas syringae, used to display R5 on the surface of bacteria.

In use, the modified E. coli can be cultured to the logarithmic stage, and IPTG with a final concentration of 1.0 ug/mL can be added to induce 3 h, and protein expression can be completed

We constructed an expression vector of R5 and its surface display carrier protein IPN fusion protein, using T7 promoter as the promoter, and induced expression, and after reviewing the literature, we selected to induce 3 h at 37 °C at a final concentration of 1.0 ug/mL in the logarithmic phase, disrupted the bacteria, and performed Western blotting experiments on the supernatant and precipitation of the cell disruption solution to verify the expression of the protein of interest.

Fig1.Bacterial holoprotein Western blotting development result

We silicified the mutant strains using the method of silicification of bacteria obtained from the literature, and the control group did the same, we collected the silicified bacteria and observed the bacteria using transmission electron microscopy to obtain the silicification effect of R5 under the silicification conditions we used.