Identification of a cDNA clone specific for the taxol synthesis phase of Taxus chinensis cells by mRNA differential display.

In order to identify genes related to Taxol biosynthesis, the mRNA differential display method was employed to compare mRNA populations from suspension cultured Taxus chinensis cells before and after beginning to produce Taxol. From a total display of about 4000 PCR products, 104 were derived from cells during the Taxol synthesis phase but not the non-Taxol synthesis phase. These products were cloned, and one such cDNA clone, named TS1, was confirmed to be specifically expressed in the Taxol synthesis phase by northern blot analysis. The length of the transcript corresponding TS1 was approximately 2.1 kb. DNA sequencing and homology search showed the sequence of TS1 contains a partial open reading frame and has no homologies with other known genes. Hence, this report demonstrated the potential of mRNA differential display for the isolation of genes specific for the period of secondary metabolite production as well as the feasibility of the approach for the identification of genes potentially related to the synthesis of secondary metabolites in higher plants.