Difference between revisions of "Part:BBa K3360003"
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The traditional fraying process is very harmful to the environment, resources and users. In essence, being old is the destruction of cellulose on the surface of jeans. We finally consider using cellulase to achieve this effect. Through cellulase washing, it can hydrolyze the cellulose and make a part of the dye fall off the fabric. Through this effect, the "worn feeling" effect that people want can be obtained. | The traditional fraying process is very harmful to the environment, resources and users. In essence, being old is the destruction of cellulose on the surface of jeans. We finally consider using cellulase to achieve this effect. Through cellulase washing, it can hydrolyze the cellulose and make a part of the dye fall off the fabric. Through this effect, the "worn feeling" effect that people want can be obtained. | ||
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<center>Fig1. Plasmid vector diagram</center> | <center>Fig1. Plasmid vector diagram</center> | ||
| + | We synthesized the above-mentioned vector and double digested it with BamHI and NotI. After DNA electrophoresis, the following map was obtained. The picture shows that after double enzyme digestion, the band 1 is lower than the band 2, and the size is about 5000bp, which is the same as our prediction, indicating that the gene transformation is successful. | ||
| − | + | [[File:Agarose-gel electrophoresis of IARI-SP-2.png |200px|center]] | |
| − | + | <center>Fig.2 Agarose-gel electrophoresis of IARI-SP-2</center> | |
<!-- Uncomment this to enable Functional Parameter display | <!-- Uncomment this to enable Functional Parameter display | ||
Revision as of 21:15, 23 October 2020
IARI-SP-2
Encoding the endo-beta-1,4-glucanase of Bacillus subtilis, the entire sequence has been optimized by E.coli codons, and the N-terminus is marked with GST to increase its solubility and reduce the formation of inclusion bodies.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1587
- 1000COMPATIBLE WITH RFC[1000]
SZU-China 2020 TEAM
The traditional fraying process is very harmful to the environment, resources and users. In essence, being old is the destruction of cellulose on the surface of jeans. We finally consider using cellulase to achieve this effect. Through cellulase washing, it can hydrolyze the cellulose and make a part of the dye fall off the fabric. Through this effect, the "worn feeling" effect that people want can be obtained.
We chose the IARI-SP-2 endoglucanase, an endo-beta-1,4-glucanase from Bacillus subtilis. We loaded it into pGEX-4T-1-H vector as shown below. Under the induction of IPTG, IARI-SP-2 will be expressed in large quantities. For IARI-SP-2, we also have carried out codon optimization to optimize its expression in E.coli. with the GST tag.
We synthesized the above-mentioned vector and double digested it with BamHI and NotI. After DNA electrophoresis, the following map was obtained. The picture shows that after double enzyme digestion, the band 1 is lower than the band 2, and the size is about 5000bp, which is the same as our prediction, indicating that the gene transformation is successful.
