MANIPULATION OF T BREVIFOLIA PHYSIOLOGY IN TISSUE CULTUR

该项目旨在广泛了解 短叶红豆杉及其近缘种的组织培养操作 用于生产紫杉醇或紫杉醇样化合物， 转化为紫杉醇。 紫杉醇是一种很有前途的抗癌药物， 供应极其短缺。 更好地理解如何操纵 在组织培养中紫杉醇的细胞合成将为 紫杉醇的有效“工厂样”生产，从而减轻 潜在的供应问题。 该项目的具体目标包括： 方法如开发紫杉醇的快速测定技术; 建立新的文化，特别是根文化; 紫杉醇自身抑制和可能的抑制机制;以及， 紫杉醇在不同树脂上分配系数的表征 和有机溶剂中。 通过这些技术，我们将探索 气相组成、激发子、原位产物去除和 细胞固定对细胞生长速率，紫杉醇生产速率， 细胞外紫杉醇与细胞内紫杉醇的比例，以及 紫杉醇相关化合物。 此外，我们将研究液体的影响， 剪切对细胞生理学的影响。 此外，新的小规模 设备将被用来测试根文化的内在能力， 增长和产品形成。

This project is directed towards a broad-based understanding of the manipulation of tissue cultures of Taxus brevifolia and related species for the production of taxol or taxol-like compounds which could be easily converted to taxol. Taxol is very promising anti-cancer agent which is in extremely short supply. A better understanding of how to manipulate cellular synthesis of taxol in tissue culture will provide the basis for efficient 'factory-like' production of taxol and thereby alleviate potential supply problems. The specific component goals of the project includes enabling methods such as development of rapid assay techniques for taxol; establishment of new cultures, particularly root cultures; evaluation of taxol self-inhibition and possible mechanisms of inhibition; and, characterization of taxol partition coefficients on a variety of resins and into organic solvents. With these techniques we will probe the effects of gas phase composition, elicitors, in situ product removal, and cell immobilization on cellular growth rates, taxol production rates, ratio of extracellular to intracellular taxol, and production of taxol-related compounds. Further, we will examine the effects of liquid shear in stirred reactors on cell physiology. Also, novel small-scale devices will be used to test the intrinsic capacity of root cultures for growth and product formation.