Part:BBa_K2317002

CbeA

CbeA-CbtA is one of the Escherichia coli TA systems.CbtA inhibits the polymerization of bacterial cytoskeletal proteins, MreB and FtsZ. The fission of bacteria will be suppressed but the growth of bacteria is normal. Cbea was found to directly interact with MreB and FtsZ, and enhance the bundling of their filamentous polymers in vitro. Besides, CbeA can relieve the toxicity of CbtA. (Figure 1.)

Figure 1. The mechanism of CbeA and CbtA(type IV TA system).

Usage and Biology

The population and growth condition can be reflected by Abs600, so through the curve of Abs600-time, the effect of CbeA and CbtA towards bacteria can be displayed.
The Abs600 values were measured within TA group and pET28a group from 0 to 16 hours and the Abs600 curves were drawn (Fiure .2). After adding IPTG, the growth rate of TA group is significantly larger than that without the addition of IPTG. However, in pER28a group, the growth rate is smaller.

Figure 2. Detoxication curve. The growth rate with addition L-Arabinose and IPTG in vector (A) or TA group (B). When OD600 value reached 0.15, L-Arabinose was added to induce the expression of CbtA. After the OD600 values remained unchanged for a while, IPTG was added simultaneous to induce the expression of CbeA. The growth curve was drawn at indicated time.

Characterization:

Through agarose gel electrophoresis, the plasmid we constructed was verified.(figure 3.) We chose DL 10000 as the marker and used BBa_K864402 as the control. Eight of the lanes contained pSB1C3-CbtA monoclones and Eight of the lanes contained pSB1C3-CbeA monoclones. After that, we sent them for sequencing and the results showed that the sequences were right.

Figure 3. The result of agarose gel electrophoresis.

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
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 116
  
• 1000
  COMPATIBLE WITH RFC[1000]