|Budget Amount *help
¥2,200,000 (Direct Cost : ¥2,200,000)
Fiscal Year 1996 : ¥800,000 (Direct Cost : ¥800,000)
Fiscal Year 1995 : ¥1,400,000 (Direct Cost : ¥1,400,000)
Exogenous expression of Rb tumor suppresser gene in cells was formerly difficult due to its intrinsic nature of inhibiting cell growth. However, the use of inducible expression system led to isolation of derivatives of NIH3T3 cell line, which will undergo complete growth arrest upon induction of RB.Incidentally, all of the information necessary for an organism is preserved as a genetic material, DNA.Therefore, realization of the information is fundamental processes of life, especially transcription step should deserve special attention. Differential display method (DD) is the most efficient and consistent method currently available for displaying transcripts present in the cells. The RB gene is speculated to exert its function by interacting with various transcriptional machinery, thus outcome of its action must be reflected in alteration of transcription. I have further modified original DD more consistent and efficient resulting in improved DD.By applying the method to RB cells, a number of genes emerged that manifested itself to be enhanced or reduced expression following RB induction. We are currently in the process of analyzing these genes. Parenthetically, we are also looking into genes involved in serum starvation, serum stimulation, contact inhibition, senescence, differentiation, and apoptosis by the same method. An extensive survey of these genes will enable us to classify genes involved in general growth arrest processes or more specialized growth arrest processes leading to discovery of more definitive and unambiguous definition of function of these genes involved in growth arrest.