Difference between revisions of "Part:BBa K2984052"

Revision as of 21:08, 18 October 2019

L1c-PsadIntron-bleRscp-ARS-YFP-Rbcs2

This vector is a part of the Chlamy-HUB-Collection. Level 1 construct containing secretion signal of arylsulfatase ARS fused to the flourescent protein mVenus. To influence the expression of mVenus, a bleomycin resistance gene is connected through a self cleaving peptide (scp) to the part. The Promoter is the PsaDintron. The construct ends with the Rbcs2 terminator.

The bleomycin resistance leads to a higher expression of the enzyme, due to its working mechanism. For every two bleomycin molecules, the bleomycin resistance gene must be expressed once. The resistance works in a ratio of 1:2 to the antibiotic (Hayes et al., 1990). By having the resistance in the same open reading frame, the enzyme is also expressed, leading to a higher expression of the enzyme (Lumbreras et al. 1998).

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal PstI site found at 2051
12
INCOMPATIBLE WITH RFC[12]
Illegal PstI site found at 2051
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 4
23
INCOMPATIBLE WITH RFC[23]
Illegal PstI site found at 2051
25
INCOMPATIBLE WITH RFC[25]
Illegal PstI site found at 2051
Illegal NgoMIV site found at 1607
Illegal NgoMIV site found at 2528
1000
COMPATIBLE WITH RFC[1000]

Characterization

colonies_total — Fig.1 - Image of a successful transformation in E.coli after ligation of the construct. The construct can then be isolated from E. coli to be transformed in C. reinhardtii

References

Lumbreras, V., Stevens, D. R., & Purton, S. (1998). Efficient foreign gene expression in Chlamydomonas reinhardtii mediated by an endogenous intron. The Plant Journal, 14(4), 441-447.
Hayes, J. D., & Wolf, C. R. (1990). Molecular mechanisms of drug resistance. Biochemical Journal, 272(2), 281.

@@ Line 6: / Line 6: @@
 This vector is a part of the <a href="https://2019.igem.org/Team:Humboldt_Berlin/Part_Collection">Chlamy-HUB-Collection</a>. Level 1 construct containing secretion signal of arylsulfatase <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984000">ARS</a> fused to the flourescent protein <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984017">mVenus</a>. To influence the expression of mVenus, a <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984045">bleomycin resistance</a> gene is connected through a self cleaving peptide (scp) to the part. The Promoter is the <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984046">PsaDintron</a>. The construct ends with the <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984018">Rbcs2 terminator</a>.
 </html>
+The bleomycin resistance leads to a higher expression of the enzyme, due to its working mechanism. For every two bleomycin molecules, the bleomycin resistance gene must be expressed once. The resistance works in a ratio of 1:2 to the antibiotic (Hayes et al., 1990). By having the resistance in the same open reading frame, the enzyme is also expressed, leading to a higher expression of the enzyme (Lumbreras et al. 1998).
 <!-- Add more about the biology of this part here
@@ Line 27: / Line 29: @@
 <partinfo>BBa_K2984052 parameters</partinfo>
 <!-- -->
+==References==
+<ol>
+<li>
+Lumbreras, V., Stevens, D. R., & Purton, S. (1998). Efficient foreign gene expression in Chlamydomonas reinhardtii mediated by an endogenous intron. The Plant Journal, 14(4), 441-447.
+</li>
+<li>
+Hayes, J. D., & Wolf, C. R. (1990). Molecular mechanisms of drug resistance. Biochemical Journal, 272(2), 281.
+</li>
+</ol>