Part:BBa_K4882000
Pmcl1 (short), a truncated version of hemolymph inducible promoter from Metarhizium anisopliae
The Pmcl1 promoter is responsible for regulating the collagen-like protein MCL1 in M. anisopliae. Pmcl1 is a hemolymph inducible promoter that turns on only in insect hemolymph (Wang et al., 2006). iGEM16_NYMU-Taipei registered the full-length Pmcl1 as BBa_K2040100. Compared to the full-length Pmcl1 (2764bp), a truncated, shorter version of Pmcl1 (1586bp) can lead to a twofold increase in downstream gene expression (Kanjo et al., 2019).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 489
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 45
- 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
M. anisopliae is an entomopathogenic fungus widely used as a biopesticide. However, it is less efficient than most chemical pesticides. Genetical engineering is the method to improve the efficiency and safety of M. anisopliae. Pmcl1 (short) can be used to control the expression of exogenous toxins and suicide switches.
Characterization
2023 Hangzhou-SDG Team characterized this part with expression of the downstream genes
KillerRed (BBa_K1184000) and SuperNova (BBa_K4882008) are red fluorescent proteins that produce reactive oxygen species (ROS) in the presence of light. They can be a part of the suicide switch for M. anisopliae when being connected after the hemolymph inducible promoter Pmcl1/Pmcl1(short).
Larvae of Galleria mellonella were killed by engineered M. anisopliae. The corpses were placed under sunshine for days until spores formed outside the insect body. Spores were collected by vertexing and the concentrations of spore suspensions were checked under a microscope.
< img class="bild" src="https://static.igem.wiki/teams/4882/wiki/part-pmcl1-short-1.png">
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