Part:BBa_K5211020
Ptet-T1AB1
-Usage and Biology
Description:
Use the RBS-10 sequence to link thetesB, phaA, and phaB genes to form theT1AB operon. （tesB derived from the E. coli K12 and phaA, phaB derived from C. necator H16
Usage:
By tandemly assembling the tesB, phaA, and phaB genes into the T1AB operon and integrating it into the pEZ15Asp plasmid, the expression of genes was regulated using the Ptet inducible promoter. To enhance the electrotransformation efficiency of the plasmid, the plasmid was transformed into the demethylated Escherichia coli trans110 competent cells, and the required plasmid was extracted again through PCR verification and culturing, ultimately obtaining the pEZ-PtT1 plasmid. Subsequently, the plasmid was electrotransformed into the ZMNPΔ0038 competent cells to construct the strain NP-PtT1 capable of producing 3-HB.
Fig.1-1 Plasmid Design of the T1AB Operon.
The fermentation performance of the recombinant strain NP-PtT1 is shown in Fig.1-2. NP-PtT1 reached a maximum 3-HB titer of 425.67±2.89 mg/L. This indicated that that our improvements can enhance the yield of 3HB.
Fig.1-2 Cell growth and 3HB production of NP-PtT1 with 1.2 μg/mL tetracycline concentrations as an inducer.
Metabolic pathway of zymomonas mobilis
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