Part:BBa_K112000

T4 holin, complete CDS, berkeley standard

T4 holin gene, with start and stop codons.

---

This part is in BBb Format. It is flanked by BamHI and BglII sites instead of XbaI and SpeI. More information about the BBb Format is available at:
[http://openwetware.org/wiki/Template:AndersonLab:BBb_Standard BBb Standard Description Page]

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]

Team NYMU-Taipei 2017: Successfully construct T4 holin into suicide mechanism

Improvement

We successfully combine T4 holin with a lactose-induced promoter (BBa_R0010), a ribosome binding site (BBa_B0034) and a double terminator (BBa_B0010 and BBa_B0012). Therefore, holin can be induced by lactose and regulate the suicide mechanism. Moreover, we construct T4 holin and T4 endolysin together as a functional suicide mechanism. Besides, in NYMU-Taipei 2017 team’s project, we also put the suicide mechanism into practice by constructing them with NrtA.

Result

Figure 1 is the gene map of T4 holin construct, pSB1C3-J04450-K112000.

Figure 1

Figure 2 is electrophoresis result of holin (from BBa_K112000) PCR product. The marker is 100bp. The length is 657bp as expected.

Figure 2

Figure 3 is electrophoresis result of holin backbone (from BBa_J04450) PCR product. The marker is 1kb. The length is 705bp as expected.

Figure 3

Figure 4 and Figure 5 are the results of suicide mechanism functional test. Both figure show that our suicide mechanism can be induced by adding lactose, and the effectiveness of suicide mechanism goes better as the concentration of lactose goes higher.

Figure 4 is the bacterium with holin-endolysin construct. As figure 3 shows, the suicide mechanism is induced immediately when lactose is added into the samples. Besides, we can see that the lactose concentration and the OD value of the bacterium with holin-endolysin construct are positively correlated, which means our suicide mechanism does work.

Figure 4

Figure 5 is the bacterium with holin-endolysin-NrtA construct. As figure 5 shows, the trend of the relative absorbance is downward as the lactose is added to induce the suicide mechanism. The concentration of lactose is also positively correlated with the declining degree of relative absorbance.

Figure 5