Part:BBa_K358019

Lysis cassette regulated by lacP

Lysis cassette[SRRz] is regulated by lactose promoter. Activating lactose promoter and expressing SRRz gene, it causes the cell lysis. So, lactose promoter must be repressed when transform this part.

To repress lactose promoter tightly, we constructed this part on low copy plasmid, pSB4K5, and transformed into KRX, not Top 10. KRX has lacI in its genome and Top 10 hasn't (see genotype[http://www.promega.com/pnotes/94/14410_27/14410_27.pdf],[http://openwetware.org/wiki/E._coli_genotypes#TOP10_.28Invitrogen.29]).

Added by THINKER_CHINA

Procedures to prove our lysis module using lac promoter:

1. Cultivate E. Coli in LB mediums at 37 degrees Celsius and 200 rpm.

2. When OD600 values equals to 0.4, add different concentrations of copper, three times for each group.

3. Measure OD600 values at 0.5h, 1h, 1.5h, 2h, 3h and 4h intervals.

Figure 1 OD600 values of different concentration of copper at various timed intervals

the graph suggested that the lysis circuit works regularly when the concentration of copper is above 10^-5 mol/L.

Figure 1 Comparison of final OD600 values after 4h、10^-5 mol/L、10^-4 mol/L (copper-sensitive promoter）

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]