Part:BBa_K4274036

PvanP*-patB_target

This is the composite part that we have designed to knock-out the patB gene in Bacillus subtilis. It starts with the promoter PvanP* (part: BBa_K4274006), a semi-synthetic promoter. Then, we have encoded the patB_target (part: BBa_ K4274007), the single guide RNA that locates the targeted gene sequence for Cas9 protein. This composite part terminates with the terminator B0015 (BBa_B0015). The parts used to knock-out patB from Bacillus subtilis, which is composed of patB_target (Part: BBa_K4274007), PvanP* (Part: BBa_K4274006).
This part collection was designed to knock-out patB gene in Bacillus subtilis, and then formed competition between Staphylococcus spp and modified Bacillus subtilis for the S-Cys-Gly-3M3SH but would not produce the malodorous molecule of 3M3SH.

Usage and Biology

We utilized the Cas9 protein to perform the knock-out of patB gene in Bacillussubtilis. patB_target (Part: BBa_K4274007) is a synthetic single guide RNA that assists the Cas9 protein to find the targeted gene sequence and perform the gene knock-out. As an extension, patB is one of the proteins in Staphylococcus. Spp. that breaks down S-Cys-Gly-3M3SH, resulting in the secretion of 3M3SH. Meanwhile, it was found that Bacillus subtilis has the identical gene sequence within its genome. Thus this part could be used to knock-out patB in B. subtilis to construct a promising bacterial strain for solving body odor problem.

Source

patB from Bacillus subtilis.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]