Reporter vector for evaluating characteristics of subject cell, assay kit, procedure and device

An assay kit for evaluating cell characteristics of a subject cell containing a reporter vector and a carrier which supports the reporter vector, the reporter vector comprising:

• (1) a first reporter gene expression unit including a promoter sequence of a cell characteristics marker, a first reporter gene existing in a downstream thereof, and a first transcription termination sequence existing in a further downstream thereof;
• (2) a second reporter gene expression unit including a promoter sequence exhibiting constitutive activity, a second reporter gene existing in a downstream thereof, a bicistronic expression sequence existing in a further downstream thereof, a replication initiation protein gene existing in a further downstream thereof, and a second transcription termination sequence existing in a further downstream thereof;
• and
• (3) a replication initiation sequence binding to a replication initiation protein synthesized from the replication initiation protein gene, thereby initiating replication of the reporter vector.

A method of evaluating cell characteristics of a subject cell, comprising:

• (a) introducing a reporter vector into the subject cell;
• (b) causing the reporter vector to replicate and expressing the first reporter gene and the second reporter gene;
• (c) detecting a first signal from the first reporter gene and a second signal from the second reporter gene;
• and
• (d) evaluating the cell characteristics of the subject cell based on the first signal and the second signal, and
• the reporter vector comprising:
• (1) a first reporter gene expression unit including a promoter sequence of a cell characteristics marker, a first reporter gene existing in a downstream thereof, and a first transcription termination sequence existing in a further downstream thereof;
• (2) a second reporter gene expression unit including a promoter sequence exhibiting constitutive activity, a second reporter gene existing in a downstream thereof, a bicistronic expression sequence existing in a further downstream thereof, a replication initiation protein gene existing in a further downstream thereof, and a second transcription termination sequence existing in a further downstream thereof;
• and
• (3) a replication initiation sequence binding to a replication initiation protein synthesized from the replication initiation protein gene, thereby initiating replication of the reporter vector.

A device for evaluating cell characteristics of a subject cell, comprising:

• a gene introduction unit which introduces a reporter vector into the subject cell;
• a thermo-regulating unit which receives a sample including the subject cell obtained in the vector introductory unit and cultures the subject cell for a desired time;
• and
• a detection unit which receives the sample from the thermo-regulating unit and detects signals produced from the first reporter gene and the second reporter gene of the subject cell included in the sample, and
• an evaluation unit which evaluates the cell characteristics of the subject cell based on a result of the detection,
• the reporter vector comprising:
• (1) a first reporter gene expression unit including a promoter sequence of a cell characteristics marker, a first reporter gene existing in a downstream thereof, and a first transcription termination sequence existing in a further downstream thereof;
• (2) a second reporter gene expression unit including a promoter sequence exhibiting constitutive activity, a second reporter gene existing in a downstream thereof, a bicistronic expression sequence existing in a further downstream thereof, a replication initiation protein gene existing in a further downstream thereof, and a second transcription termination sequence existing in a further downstream thereof;
• and
• (3) a replication initiation sequence binding to a replication initiation protein synthesized from the replication initiation protein gene, thereby initiating replication of the reporter vector.