Difference between revisions of "Part:BBa K3114027"

 
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===Usage and Biology===
 
===Usage and Biology===
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Pheophytinase is a chloroplast-located hydrolase which dephytylates the pigment pheophytin during the chlorophyll degradation pathway in plants and algae (Schelbert et al., 2009). Pheophytinase catalyzes the reaction removing the phytyl chain from pheophytin resulting in the formation of pheophorbide a. In several plant species, it has been shown that absence of pheophytinase results in a stay-green phenotype (Guyer et al., 2018).
  
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BBa_K3114024 is a part consisting of multiple components of iGEM Calgary's 2019 Biobrick submission kit as well as other widely used Biobricks. The coding region consists of the PPH gene which encodes for pheopytinase[https://parts.igem.org/Part:BBa_K3114011 (BBa_K3114011)].A T7 promoter[https://parts.igem.org/Part:BBa_I719005 (BBa_I719005)]was used in conjunction with a strong ribosome binding site[https://parts.igem.org/Part:BBa_B0030 (BBa_B0030)].Following the coding region is iGEM Calgary's universal spacer sequence and 6XHis tag site[https://parts.igem.org/Part:BBa_K3114014 (BBa_K3114014)].The universal spacer was utilized in an attempt to mitigate any adverse effects that the his tag may have on the protein's function, as hypothesized in Meguro et al. 2011.
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<span class='h3bb'>Sequence and Features</span>
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===Sequences and Features===
 
<partinfo>BBa_K3114027 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3114027 SequenceAndFeatures</partinfo>
  
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===References===
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Luzia Guyer, Kathrin Salinger, Undine Krügel, Stefan Hörtensteiner, Catalytic and structural properties of pheophytinase, the phytol esterase involved in chlorophyll breakdown, Journal of Experimental Botany, Volume 69, Issue 4, 6 February 2018, Pages 879–889, https://doi.org/10.1093/jxb/erx326.
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Meguro, M., Ito, H., Takabayashi, A., Tanaka, R., & Tanaka, A. (2011). Identification of the 7-hydroxymethyl chlorophyll a reductase of the chlorophyll cycle in Arabidopsis. The Plant Cell, 23(9), 3442-3453.
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Schelbert, S., Aubry, S., Burla, B., Agne, B., Kessler, F., Krupinska, K., & Hörtensteiner, S. (2009). Pheophytin pheophorbide hydrolase (pheophytinase) is involved in chlorophyll breakdown during leaf senescence in Arabidopsis. The Plant Cell, 21(3), 767-785.
  
 
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Revision as of 02:29, 20 October 2019


Pheophytinase (PPH)

Usage and Biology

Pheophytinase is a chloroplast-located hydrolase which dephytylates the pigment pheophytin during the chlorophyll degradation pathway in plants and algae (Schelbert et al., 2009). Pheophytinase catalyzes the reaction removing the phytyl chain from pheophytin resulting in the formation of pheophorbide a. In several plant species, it has been shown that absence of pheophytinase results in a stay-green phenotype (Guyer et al., 2018).

BBa_K3114024 is a part consisting of multiple components of iGEM Calgary's 2019 Biobrick submission kit as well as other widely used Biobricks. The coding region consists of the PPH gene which encodes for pheopytinase(BBa_K3114011).A T7 promoter(BBa_I719005)was used in conjunction with a strong ribosome binding site(BBa_B0030).Following the coding region is iGEM Calgary's universal spacer sequence and 6XHis tag site(BBa_K3114014).The universal spacer was utilized in an attempt to mitigate any adverse effects that the his tag may have on the protein's function, as hypothesized in Meguro et al. 2011.

Sequences and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1579
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 267
    Illegal AgeI site found at 558
  • 1000
    COMPATIBLE WITH RFC[1000]

References

Luzia Guyer, Kathrin Salinger, Undine Krügel, Stefan Hörtensteiner, Catalytic and structural properties of pheophytinase, the phytol esterase involved in chlorophyll breakdown, Journal of Experimental Botany, Volume 69, Issue 4, 6 February 2018, Pages 879–889, https://doi.org/10.1093/jxb/erx326.

Meguro, M., Ito, H., Takabayashi, A., Tanaka, R., & Tanaka, A. (2011). Identification of the 7-hydroxymethyl chlorophyll a reductase of the chlorophyll cycle in Arabidopsis. The Plant Cell, 23(9), 3442-3453.

Schelbert, S., Aubry, S., Burla, B., Agne, B., Kessler, F., Krupinska, K., & Hörtensteiner, S. (2009). Pheophytin pheophorbide hydrolase (pheophytinase) is involved in chlorophyll breakdown during leaf senescence in Arabidopsis. The Plant Cell, 21(3), 767-785.