Difference between revisions of "Part:BBa K3771079"
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<br><b style="font-size:1.3rem">Usage</b> | <br><b style="font-size:1.3rem">Usage</b> | ||
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| − | + | <br>JJU10 enzyme was used in vitro testing of taurine production. The sequence for JJU10 enzyme and trc promoter were ligated and transformed into E. coli to calculate taurine production using high-performance liquid chromatography (HPLC). A 6xHis-tag is added to the C-terminal of the JJU10 protein, allowing for confirmation of JJU10 expression by western blot using the anti-6xHis antibody. <br> | |
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<br><b style="font-size:1.3rem">Characterization</b> | <br><b style="font-size:1.3rem">Characterization</b> | ||
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<p align="center">Fig. 6 Confirmation of protein expression of JJU. | <p align="center">Fig. 6 Confirmation of protein expression of JJU. | ||
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<p align="center">Fig. 8 Confirmation of protein expression of JJU. | <p align="center">Fig. 8 Confirmation of protein expression of JJU. | ||
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<br><b style="font-size:1.3rem">References</b> | <br><b style="font-size:1.3rem">References</b> | ||
| − | <br> | + | <br>BRENDA - Information on EC 4.4.1.10 - cysteine lyase. Brenda-enzymes.org. Published 2021. Accessed October 6, 2021. https://www.brenda-enzymes.org/enzyme.php?ecno=4.4.1.10<br> |
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===Usage and Biology=== | ===Usage and Biology=== | ||
Revision as of 03:30, 19 October 2021
T7-JJU
Description
PT7-jju is a composite part consisting of the T7 promoter and the cs sequences. This part was used in in vivo testing of taurine production. The sequence for jju and T7 promoter were ligated and transformed into E. coli to calculate taurine production using high-performance liquid chromatography (HPLC).
Biology
T7 promoter constitutively facilitates the expression of JJU enzyme.
Usage
JJU10 enzyme was used in vitro testing of taurine production. The sequence for JJU10 enzyme and trc promoter were ligated and transformed into E. coli to calculate taurine production using high-performance liquid chromatography (HPLC). A 6xHis-tag is added to the C-terminal of the JJU10 protein, allowing for confirmation of JJU10 expression by western blot using the anti-6xHis antibody.
Characterization
Fig 1. Confirmation of jju fragment by PCR. M: Marker; Lane 1: jju (1180bp)
Fig 2. Confirmation of pET28a fragment by digestion. M: Marker; Lane 1: pET28a (~5280 bp)
Fig. 3 Transformation/JJU in DH5α
Fig. 4 Confirmation of pET28a-T7-JJU by digestion. M: Marker; Lane 1: Colony of pET28a-T7-JJU (~6466 bp)
Fig. 5 Transform into BL21(DE3)
Fig. 6 Confirmation of protein expression of JJU. M: Marker; Lane1: whole cell of JJU in BL21(DE3); Lane2: soluble protein of JJUBL21(DE3) (~43 kDa)
Fig. 7 Transform into BD7G
Fig. 8 Confirmation of protein expression of JJU. M: Marker; Lane1: whole cell of JJU in BD7G; Lane2: soluble protein of JJU in BD7G(~43 kDa)
References
BRENDA - Information on EC 4.4.1.10 - cysteine lyase. Brenda-enzymes.org. Published 2021. Accessed October 6, 2021. https://www.brenda-enzymes.org/enzyme.php?ecno=4.4.1.10
Sequence and Features