| 摘要 |
A sequential gene expression network is provided to be used for gene therapy or be applied to a biotechnology field by allowing a protein to be expressed at a specific time or allowing various proteins to be expressed sequentially. And a method for tagging cells is provided to be used for performing a high-throughput cell-based assay without a cell chip and various equipments. A method for sequentially expressing a target protein comprises the steps of: (a) preparing a gene construct consisting of a polynucleotide encoding an (n)th target protein(Pn) and an (n)th transcription factor(TFn) operably linked to a transcription control factor of which the expression is induced by an inducer and a polynucleotide encoding an antisense RNAn complementarily binding to a gene of an (n+1)th target protein(Pn+1) and an (n+1)th transcription factor(TFn+1); (b) preparing a gene construct consisting of a polynucleotide encoding the (n+1)th target protein(Pn+1) and the (n+1)th transcription factor(TFn+1) and a polynucleotide encoding an antisense RNAn+1 complementarily binding to a gene of an (n+2)th target protein(Pn+2) and an (n+2)th transcription factor(TFn+2); (c) transforming a host cell using the gene constructs prepared by the steps(a) and (b); and (d) treating the transformed host cell with an inducer. A sequential gene expression system comprises gene sets respectively encoding the Pn, TFn and RNAn. A method for tagging cells comprises the steps of: (a) linking an (n)th target protein to a labeling protein and then expressing it; and (b) sequentially expressing the labeling protein in accordance with the method above to respectively distinguish cells. |
