Difference between revisions of "Part:BBa K3114026"
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===Usage and Biology=== | ===Usage and Biology=== | ||
The conversion of chlorophyll a to pheophytin a is the first step in the natural chlorophyll degradation pathway. The enzyme Magnesium Dechelatase, encoded by the SGR gene, catalyzes this reaction by extracting magnesium (Mg2+) from chlorophyll, along with cofactors 2H+ (Shimoda et al. 2016). | The conversion of chlorophyll a to pheophytin a is the first step in the natural chlorophyll degradation pathway. The enzyme Magnesium Dechelatase, encoded by the SGR gene, catalyzes this reaction by extracting magnesium (Mg2+) from chlorophyll, along with cofactors 2H+ (Shimoda et al. 2016). | ||
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BBa_K3114026 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 SGR gene which encodes for the enzyme Magnesium Decheletase[https://parts.igem.org/Part:BBa_K3114010 (BBa_K3114010)].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. | BBa_K3114026 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 SGR gene which encodes for the enzyme Magnesium Decheletase[https://parts.igem.org/Part:BBa_K3114010 (BBa_K3114010)].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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| + | This part can be used for IPTG-inducible expression and secretion of the protein Chlorophyll B Reductase. This circuit contains the DsbA signal peptide. The DsbA signal peptide is a 19-amino acid sequence which targets fused proteins to the Sec secretion pathway in E. coli (Schierle et al., 2003). The fused protein is not folded until it is secreted to the periplasm. The signal peptide is cleaved after residue 19 by signal peptidase after secretion (recognition sequence: Ala-X-Ala). | ||
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===References=== | ===References=== | ||
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. | 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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| + | Schierle, C. F., Berkmen, M., Huber, D., Kumamoto, C., Boyd, D., & Beckwith, J. (2003). The DsbA signal sequence directs efficient, cotranslational export of passenger proteins to the Escherichia coli periplasm via the signal recognition particle pathway. Journal of Bacteriology, 185(19), 5706–5713. https://doi.org/10.1128/JB.185.19.5706-5713.2003 | ||
Shimoda, Y., Ito, H., & Tanaka, A. (2016). Arabidopsis STAY-GREEN, Mendel’s green cotyledon gene, encodes magnesium-dechelatase. The Plant Cell, 28(9), 2147-2160. | Shimoda, Y., Ito, H., & Tanaka, A. (2016). Arabidopsis STAY-GREEN, Mendel’s green cotyledon gene, encodes magnesium-dechelatase. The Plant Cell, 28(9), 2147-2160. | ||
Latest revision as of 11:07, 21 October 2019
Magnesium Dechelatase Stay Green (SGR) inducible circuit
Usage and Biology
The conversion of chlorophyll a to pheophytin a is the first step in the natural chlorophyll degradation pathway. The enzyme Magnesium Dechelatase, encoded by the SGR gene, catalyzes this reaction by extracting magnesium (Mg2+) from chlorophyll, along with cofactors 2H+ (Shimoda et al. 2016).
BBa_K3114026 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 SGR gene which encodes for the enzyme Magnesium Decheletase(BBa_K3114010).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.
This part can be used for IPTG-inducible expression and secretion of the protein Chlorophyll B Reductase. This circuit contains the DsbA signal peptide. The DsbA signal peptide is a 19-amino acid sequence which targets fused proteins to the Sec secretion pathway in E. coli (Schierle et al., 2003). The fused protein is not folded until it is secreted to the periplasm. The signal peptide is cleaved after residue 19 by signal peptidase after secretion (recognition sequence: Ala-X-Ala).
Sequences and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 2131
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1554
- 1000COMPATIBLE WITH RFC[1000]
References
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.
Schierle, C. F., Berkmen, M., Huber, D., Kumamoto, C., Boyd, D., & Beckwith, J. (2003). The DsbA signal sequence directs efficient, cotranslational export of passenger proteins to the Escherichia coli periplasm via the signal recognition particle pathway. Journal of Bacteriology, 185(19), 5706–5713. https://doi.org/10.1128/JB.185.19.5706-5713.2003
Shimoda, Y., Ito, H., & Tanaka, A. (2016). Arabidopsis STAY-GREEN, Mendel’s green cotyledon gene, encodes magnesium-dechelatase. The Plant Cell, 28(9), 2147-2160.