Hu Guobin, Fan Tingjun, Mei Xingguo
College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China.
Nat Prod Res. 2004 Aug;18(4):365-71. doi: 10.1080/14786410310001630573.
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.
为了鉴定与紫杉醇生物合成相关的基因，采用mRNA差异显示方法比较了中国红豆杉悬浮培养细胞在开始产生紫杉醇前后的mRNA群体。在约4000个PCR产物的总显示中，有104个来自紫杉醇合成阶段的细胞，而非紫杉醇合成阶段的细胞中没有。这些产物被克隆，通过Northern印迹分析证实其中一个名为TS1的cDNA克隆在紫杉醇合成阶段特异性表达。与TS1对应的转录本长度约为2.1 kb。DNA测序和同源性搜索表明，TS1的序列包含一个部分开放阅读框，与其他已知基因无同源性。因此，本报告证明了mRNA差异显示在分离次生代谢产物产生时期特异性基因方面的潜力，以及该方法在鉴定高等植物中潜在与次生代谢产物合成相关基因方面的可行性。
