Difference between revisions of "Team:Mingdao/Basic Part"

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<p>To express CYP2E1 gene in Bacillus subtilis, we transfer the DNA fragments of PliaI-RBS-CYP2E1-Tr to pBS0E vector and transformed Bacillus subtilis 168 with the resulting plasmids.
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Revision as of 12:02, 20 October 2019

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Best Basic Part

CYP2E1 efficiency measure

Emerson reagent

Protocol

Reference

Best Basic parts

Part name:CYP2E1, rabbit cytochrome P450 2E1 ( optimized for Bacillus preferred codon)

Part cat. no.:BBa_K2932000

Biological function

Benzene oxidation

Phenol is a product from benzene oxidation which is catalyzed by CYP2E1.

Gene cloning and protein expression

We optimized the gene codon of rabbit cytochrome P450 2E1 (CYP2E1) based on Bacillus subtilis codon usage preference and synthesized the DNA fragment by Twist Bioscience. Then, DNA fragments were amplified by PCR and assembled with a terminator, followed by inserting to PliaI-RBS/pSB1C3.

Construct of PliaI-RBS-CYP2E1-Tr/pSB1C3



To express CYP2E1 gene in Bacillus subtilis, we transfer the DNA fragments of PliaI-RBS-CYP2E1-Tr to pBS0E vector and transformed Bacillus subtilis 168 with the resulting plasmids.