Team:Nankai/Basic Part
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Basic Parts
BBa_K1628001 (Pbca)
Promoter Pbca is the original promoter of pgsBCA operon. According to our promoter strength assay (showed in Figure 1), the strength of Pbca is very weak.
BBa_K1628002 (Pxyl)
Promoter Pxyl is a promoter of xylose operon regulate by repressor XylR. Promoter Pxyl together with promoter PlacI, repressor LacI (BBa_K1628201), promoter Pgrac (BBa_K1628202) and repressor XylR (BBa_K1628203) formed a metabolic toggle switch. We used this device to regulate the expression of odhAB genes in B. amyloliquefaciens NK-1 (showed in Figure 2). Without IPTG, the promoter Pgrac is inhibited by suppressor LacI and the supreessor XylR will not synthesized, thus the promoter Pxyl is active and odhAB genes are expressed. When IPTG is added, the xylR gene is expressed and the suppressor XylR is synthesized thereafter inhibited the expression of odhAB genes.
BBa_K1628003 (BJ27UP )
Promoter BJ27UP is an artificially synthesized promoter. According to our promoter strength assay in B. amyloliquefaciens NK-1 (showed in Figure 1), the strength of BJ27UP is stronger than Pbcabut is still too weak compared with other promoters.
BBa_K1628004 (C2up)
Promoter C2up is an artificially synthesized promoter. According to our promoter strength assay in B. amyloliquefaciens NK-1 (showed in Figure 1), C2up is the strongest promoters we used in our project.
BBa_K1628005 (A2up)
Promoter A2cup is an artificially synthesized promoter. According to our promoter strength assay in B. amyloliquefaciens NK-1 (showed in Figure 1), A2up is a very strong promoter in B. amyloliquefaciens NK-1.
BBa_K1628006 (P43)
Promoter P43 is a strong promoter in Bacillus subtilis 168. According to our promoter strength assay in (showed in Figure 1), P43 is a weak promoter in B. amyloliquefaciens NK-1.
BBa_K1628007 (PamyA)
Promoter PamyA is a strong promoter in Bacillus amyloliquefaciens LL3. According to our promoter strength assay in (showed in Figure 1), PamyAis the second strongest promoter in our project.
BBa_K1628101 (pgsB)
pgsB is a gene responsible for γ-PGA synthesis in pgsBCA operon. 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).
BBa_K1628102 (pgsAC)
pgsC and pgsA are two genes in pgsBCA operon. Protein PgsBCA is a membrane protein responsible for the synthesis of γ-PGA. Subunit PgsC is responsible for glutamic acid’s polymerization and subunit PgsA is responsible for the secretion of γ-PGA (showed in Figure 3)
Figure 1. Promoter strength assay in Bacillus amyloliquefaciens NK-1.
Figure 3. The synthetic pathway of γ-PGA
