Part:BBa_K3191301
pBAD + GarKS
Garvicin KS (GarKS) is a leaderless bacteriocin originally isolated from Lactococcus garvieae KS1546. GarKS possesses a large spectrum of antimicrobial activity against Gram-positive genera such as Staphylococcus, Listeria, Bacillus, and Enterococcus. GarKS is composed of three peptides that are 32-34 amino acids in length. Each peptide is encoded by one of three genes located next to one another: gakA, gakB, and gakC.
This part contains gakA, gakB, and gakC for the production of the complete GarKS bacteriocin. These genes are under the influence of a pBAD promoter and separated by ribosome binding sites. Both T1 and T7TE terminators exist at the end of the sequence.
Figure 1. Protein concentration obtained from cell supernatant measured in Bradford assay.
Figure 2. Protein sample measured in Tris-Tricine gel. The red bar indicates the expected location of garvicin KS.
The concentrations of protein collected from a Bradford assay can be seen in figure 1. The negative control, pSB3K3, had the lowest noted concentration of 0.335 mg/mL. This indicates the basal level of protein production in the cells. The culture containing the garvicin KS gene displayed a protein concentration of 0.528 mg/mL, indicating an increase in the concentration of protein produced and excreted from the cell. The culture expressing garvicin with the NSP4 secretion peptide demonstrated a concentration of >1.044 mg/mL. This suggests that the inclusion of the secretion peptide successfully increased protein secretion.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 125
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 65
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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