Recombinant Expression of Isopentenyl Monophosphate Kinase
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Corresponding Organization : North Carolina State University
Other organizations : Amyris (United States), University of Pittsburgh
Variable analysis
- Codon optimization of isopentenyl monophosphate kinase (IPK) gene from Thermoplasma acidophilum DSM 1728
- Successful cloning of the codon-optimized ipk gene into the pET28a vector
- Verification of the ipk gene sequence
- Thermoplasma acidophilum DSM 1728 as the source of the ipk gene
- Use of Phire Hot Start II polymerase for PCR amplification
- Use of NdeI and XhoI restriction sites for cloning the ipk gene into pET28a
- Use of T4 ligase for ligation of the digested PCR product and pET28a vector
- Transformation into E. coli DH5α strain
- Selection of transformants on LB agar plates containing 50 μg/mL kanamycin
- Not specified
- Not specified
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