Team:Nankai/Basic Part
Basic Parts
Promoters
According to our promoter strength assay (showed in Figure 1), BBa_K1628001 (Pbca), BBa_K1628003 (BJ27UP), BBa_K1628004 (C2up), BBa_K1628005 (A2up), BBa_K1628006 (P43), and BBa_K1628007 (PamyA) show different properties. Promoter Pbca is the original promoter of pgsBCA operon.Promoter BJ27UP C2up and A2up are an artificially synthesized promoters. Promoter P43 is a strong promoter in Bacillus subtilis 168. Promoter PamyA is a strong promoter in Bacillus amyloliquefaciens LL3.
Judging from the promoter strength assay, BBa_K1628004 (C2up) is the strongest promoters we used in our project and BBa_K1628007 (PamyA) is the second strongest promoter in our project. The strength of BBa_K1628001 (Pbca) is very weak. While the strength of BBa_K1628003 (BJ27UP) is stronger than Pbca, it is still too weak compared with other promoters. What's more, BBa_K1628005 (A2up) is a very strong promoter and BBa_K1628006 (P43) is a weak promoter.
Figure 1. Promoter strength assay in Bacillus amyloliquefaciens NK-1.
Coding genes
BBa_K1628101 (pgsB) and BBa_K1628102 (pgsAC) are genes in pgsBCA operon. pgsB is a gene responsible for γ-PGA synthesis.Protein PgsBCA is a membrane protein and subunit PgsB’s main function is gathering substrate glutamic acid for γ-PGA synthesis (showed in Figure 3). Subunit PgsC is responsible for glutamic acid’s polymerization and subunit PgsA is responsible for the secretion of γ-PGA.
Figure 3. The synthetic pathway of γ-PGA
