MANIPULATION OF T BREVIFOLIA PHYSIOLOGY IN TISSUE CULTUR

本项目旨在广泛地了解 短叶红豆杉及其近缘种组织培养的操作 用于生产紫杉醇或紫杉醇类化合物，这些化合物很容易 转化为紫杉醇。紫杉醇是一种很有前途的抗癌药物 供不应求。更好地理解如何操纵 在组织培养中细胞合成紫杉醇将为 高效的工厂化生产紫杉醇，从而减轻 潜在的供应问题。 该项目的具体组件目标包括启用 方法如发展紫杉醇快速检测技术； 新培养物，特别是根培养物的建立.评价 紫杉醇的自我抑制作用及可能的抑制机制； 紫杉醇在多种树脂上分配系数的表征 转化为有机溶剂。使用这些技术，我们将探索 气相组成、激发子、原位产物去除和 细胞固定化对细胞生长率、紫杉醇产量的影响 胞外紫杉醇与胞内紫杉醇的比例以及紫杉醇的产量 紫杉醇相关化合物。此外，我们将研究液体的影响 搅拌反应器中的剪切力对细胞生理的影响。此外，新奇的小规模 将使用设备来测试根培养物的内在容量 生长和产品形成。

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.