Difference between revisions of "Part:BBa K358019"

Latest revision as of 11:53, 9 October 2022

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]

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K358019 short</partinfo>
-Lysis cassette[SRRz] is regulated by lactose promoter.  Activate lactose promoter and express SRRz gene, it causes the cell lysis.
+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:
+.  Cultivate E. Coli in LB mediums at 37 degrees Celsius and 200 rpm.
+.  When OD600 values equals to 0.4, add different concentrations of copper, three times for each group.
+.  Measure OD600 values at 0.5h, 1h, 1.5h, 2h, 3h and 4h intervals.
+[[Image: T--THINKER_CHINA--THINKER_CHINA4.jpg | thumb | center | 300px |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.
+[[Image: T--THINKER_CHINA--THINKER_CHINA5.jpg | thumb | center | 300px |Figure 1 Comparison of final OD600 values after 4h、10^-5 mol/L、10^-4 mol/L (copper-sensitive promoter） ]]
 <!-- Add more about the biology of this part here