Difference between revisions of "Part:BBa K1897013:Design"

Latest revision as of 13:24, 15 October 2016

Listeriolysin O

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 1482
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 90
1000
COMPATIBLE WITH RFC[1000]

Design Notes

There is a HA tag available for characterisation of the protein produced via western blot. Note that the stop codon for LLO CDS BBa_K1897012 is shifted to after the HA tag. Also, the transcriptional terminators rrnBT1 (from BBa_B0010) + BBa_B0012 was derived from BBa_B0015, with the first 8 base pairs removed.

Source

Listeriolysin O (LLO) is derived from Listeria monocytogenes and is initially obtained from BBa_K177026. However, after sequencing, the results obtained for sequencing of BBa_K177026 did not appear to match that given on the page. We went on to synthesize new LLO (for BBa_K1897013) with the results obtained from sequencing, and also added a HA tag.

References

Kayal, S., & Charbit, A. (2006). Listeriolysin O: a key protein of Listeria monocytogenes with multiple functions. FEMS microbiology reviews, 30(4), 514-529.

Vadia, S., Arnett, E., Haghighat, A. C., Wilson-Kubalek, E. M., Tweten, R. K., & Seveau, S. (2011). The pore-forming toxin listeriolysin O mediates a novel entry pathway of L. monocytogenes into human hepatocytes. PLoS Pathog, 7(11), e1002356.

@@ Line 7: / Line 7: @@
 ===Design Notes===
-There is a HA tag available for characterisation of the protein produced via western blot. Note that the stop codon for LLO CDS <partinfo>BBa_K1897012</partinfo> is shifted to after the HA tag.
+There is a HA tag available for characterisation of the protein produced via western blot. Note that the stop codon for LLO CDS <partinfo>BBa_K1897012</partinfo> is shifted to after the HA tag. Also, the transcriptional terminators rrnBT1 (from <partinfo>BBa_B0010</partinfo>) + <partinfo>BBa_B0012</partinfo> was derived from <partinfo>BBa_B0015</partinfo>, with the first 8 base pairs removed.
@@ Line 13: / Line 13: @@
 ===Source===
-Listeriolysin O (LLO) is derived from <i>Listeria monocytogenes</i> and is _______.
+Listeriolysin O (LLO) is derived from <i>Listeria monocytogenes</i> and is initially obtained from <partinfo>BBa_K177026</partinfo>. However, after sequencing, the results obtained for sequencing of <partinfo>BBa_K177026</partinfo> did not appear to match that given on the page. We went on to synthesize new LLO (for <partinfo>BBa_K1897013</partinfo>) with the results obtained from sequencing, and also added a HA tag.
 ===References===
+Kayal, S., & Charbit, A. (2006). Listeriolysin O: a key protein of Listeria monocytogenes with multiple functions. <i>FEMS microbiology reviews, 30</i>(4), 514-529.
+Vadia, S., Arnett, E., Haghighat, A. C., Wilson-Kubalek, E. M., Tweten, R. K., & Seveau, S. (2011). The pore-forming toxin listeriolysin O mediates a novel entry pathway of L. monocytogenes into human hepatocytes. <i>PLoS Pathog, 7</i>(11), e1002356.