Difference between revisions of "Part:BBa K3779009:Experience"

Revision as of 12:48, 10 October 2021

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Applications of BBa_K3779009

① Codon optimization</br> The original target gene BBa_K3779022 was codon optimized according to pichia pastoris preference, and the optimized codon SS-bgly (BBa_K3779001) was used for plasmid construction to improve the activity of our enzyme. ②the signal peptide</br> We added the signal peptide α-factor (BBa_K3779002) to our plasmid, which contributed to the extracellular expression of our target protein in pichia pastoris. ③ AOX1 promoter</br> We used methanol-induced promoters and designed 5 'and 3' primers BBa_K3779000 and BBa_K3779003. Integration of foreign genes ④ Integration of foreign genes</br> Linearized plasmids were introduced into p. Pastoris genome for homologous integration by electrotransformation. This method is simple and efficient in practice.

The recombinant expression strain of pichia pastoris β -glycosidase was constructed, as shown in figure. <img src="" style="width:200px;height:200px;"></a> Results As shown in the figure, there was a target band of about 2000 bp in line with the theoretical size, while the blank control GS115-pPIC9K only had a band of 500 bp, proving that the recombinant strain was successfully constructed. （3）链接

User Reviews

UNIQbbe9250e9eb28b1a-partinfo-00000000-QINU UNIQbbe9250e9eb28b1a-partinfo-00000001-QINU

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 ===Applications of BBa_K3779009===
-① Codon optimization
+<div>
-The original target gene BBa_K3779022 was codon optimized according to pichia pastoris preference, and the optimized codon SS-bgly (BBa_K3779001) was used for plasmid construction to improve the activity of our enzyme.
+<b>① Codon optimization</b></br>
-②the signal peptide
+&nbsp;&nbsp;The original target gene BBa_K3779022 was codon optimized according to pichia pastoris preference, and the optimized codon SS-bgly (BBa_K3779001) was used for plasmid construction to improve the activity of our enzyme.
-We added the signal peptide α-factor (BBa_K3779002) to our plasmid, which contributed to the extracellular expression of our target protein in pichia pastoris.
+<b>②the signal peptide</b></br>
-③ AOX1 promoter
+&nbsp;&nbsp;We added the signal peptide α-factor (BBa_K3779002) to our plasmid, which contributed to the extracellular expression of our target protein in pichia pastoris.
-We used methanol-induced promoters and designed 5 'and 3' primers BBa_K3779000 and BBa_K3779003.
+<b>③ AOX1 promoter</b></br>
+&nbsp;&nbsp;We used methanol-induced promoters and designed 5 'and 3' primers BBa_K3779000 and BBa_K3779003.
 Integration of foreign genes
-④ Integration of foreign genes
+<b>④ Integration of foreign genes</b></br>
-Linearized plasmids were introduced into p. Pastoris genome for homologous integration by electrotransformation. This method is simple and efficient in practice.
+&nbsp;&nbsp;Linearized plasmids were introduced into p. Pastoris genome for homologous integration by electrotransformation. This method is simple and efficient in practice.
+</div>
+<div>
+&nbsp;&nbsp;The recombinant expression strain of pichia pastoris β -glycosidase was constructed, as shown in figure.
+<img src="https://2021.igem.org/wiki/images/2/29/T--NWU-CHINA-B--pPIC9K-SS-bgly.png" style="width:200px;height:200px;"></a>
+Results As shown in the figure, there was a target band of about 2000 bp in line with the theoretical size, while the blank control GS115-pPIC9K only had a band of 500 bp, proving that the recombinant strain was successfully constructed.
+（3）链接
+</div>
 ===User Reviews===