Difference between revisions of "Part:BBa K2350021"

Revision as of 02:30, 28 October 2017

R0010-B0034-Holin-B0010-B0012-J23106-B0034-Endolysin-B0010-B0012-J23118-B0034-NrtA-B0015

Holin-Endolysin-NrtA

　　In this part, we combine holin (BBa_K2350019), endolysin (BBa_K2350020), and NrtA (BBa_K2350004). NrtA protein can stick to periplasmic membrane through a flexible linker to capture nitrite or nitrate in the periplasm. Therefore, we make use of NrtA in nitrogen starvation in our 2017 NYMU project. After we reach our goal, we need to kill E.coli to prevent contamination. When the lactose is added into the environment, the suicide mechanism, holin and endolyin, is induced. For more detail information, please see in BBa_2350004 for NrtA part, BBa_K2350018 for endolysin part, BBa_K2350019 for holin part, and BBa_K2350020 for endolysin-holin part.

Promoter, RBS, and Terminator

　　For more details of the connected promoter, RBS, and terminator, you can see in BBa_2350004 for NrtA part, BBa_K2350018 for endolysin part, and BBa_K2350019 for holin part.

Result

　　Figure 1 is restriction enzyme check electrophoresis result of Holin-Endolysin-NrtA construct. We use XmaI to digest sample 1~3. The result shows that 2 and 3 are right. M represents 1 kb marker.

Figure 1

Functional Test

　　As figure 2 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 2