Part:BBa_K1992005

Tar native RBS expression system (promoter+RBS+coding+terminator)

introduction Tar chemoreceptor is one of four native chemoreceptors of E.coli, it is mediate chemotaxis response towards aspartate and away from Ni and Co (מקור). This device allows the expression of a functionally native Tar chemoreceptor.

usage and biology This device is very similar to our K1992004 device the only difference is the RBS, here the expression system is using the native RBS of the Tar receptor as found in the E.coli genome (K1992000).

expriments and results In order to test the device it was cloned to chemoreceptor free strain (UU1250) and tested for chemotaxis ability using swarming assay. As can be seen in figure 1 the cells expressing the Tar chemoreceptor create a halo indicating a functional chemotaxis result, which compared to the Tar expression system using a strong RBS(B0034).

Figure 1

Fig1. (a) Tar expression in UU1250 strain cloned with K1992004 expretion system - strong RBS. (b) Tar expression in UU1250 strain cloned with K1992004 expretion system - Tar native RBS

The part was successfully sequenced (fig2) ensuring the present of the promoter, RBS, Tar receptor and terminator.

Figure 2

Fig 2. (a) Sequencing chromatogram of the K1992005 BioBrick- the RBS is marked. (b) The alignment of the sequencing results in compared to the designed sequence - alignment was done using snapgene software.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1355
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 184