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.