Plasmid Maps for a Nuclear Transformation Vector in Chlamydomonas reinhardtii for the Expression and Secretion of the Plastic-Degrading Enzyme (PHL7)

pJP32PHL7 Vector:

• Size: 5692 bp

• Key Features:

  • HSP70 Promoter: A heat shock protein promoter fused with the rbcS2 promoter to drive expression of downstream genes.
  • Ble Resistance Gene: Confers resistance to bleomycin, useful for selection in Chlamydomonas reinhardtii.
  • PHL7 Gene: Encodes the plastic-degrading enzyme PHL7, inserted downstream of the F2A site for expression in the host.
  • Intron Sequences: Contains multiple rbcS2 introns for enhancing expression in Chlamydomonas.
  • Selectable Marker (AmpR): Confers ampicillin resistance for selection in E. coli.
  • Replication Origin: Includes ori and F1 ori for replication in E. coli.

   

• Applications: This vector is designed for nuclear transformation in Chlamydomonas reinhardtii, enabling the expression and secretion of the plastic-degrading enzyme (PHL7) under the control of a hybrid HSP70rbcS2 promoter.

pJP32PHL7dg Vector:

• Size: 5692 bp

• Key Features:

  • HSP70 Promoter: Retains the HSP70 and rbcS2 fusion promoter for gene expression.
  • LacZ Alpha Fragment: Includes a LacZ alpha fragment for blue/white screening.
  • PHL7 Gene: Encodes the plastic-degrading enzyme PHL7, linked downstream of the F2A site, allowing for expression in the host.
  • Ble Resistance Gene: Also confers bleomycin resistance for selection in Chlamydomonas.
  • Selectable Marker (AmpR): Confers ampicillin resistance for selection in E. coli.
  • Intron Sequences: Contains rbcS2 introns for optimizing gene expression in the host organism.

   

• Applications: The pJP32PHL7dg vector is similarly designed for nuclear transformation in Chlamydomonas reinhardtii. It also facilitates the expression and secretion of the plastic-degrading enzyme PHL7, driven by the hybrid HSP70rbcS2 promoter, but without glycosilation sites.