IDENTIFICATION OF DRUG TARGETS THROUGH GENETICS

我们工作的总体目标是利用现代遗传学来识别 新药治疗前景看好的新药靶点(S) 艾滋病患者中的弓形虫感染。我们的方法一直是开发一种 基因图谱和适当的方法，使我们能够识别 并克隆突变形式的基因，这些基因可以导致对给定的 毒品。在未来，我们计划继续并扩大这种遗传 阿托瓦酮、克林霉素等耐药基因的定位方法 有希望的新药，因为它们是可用的。要做到这一点，我们首先 扩展我们的限制片段长度多态(RFLP)探针集 包括代表新的细胞器基因组的那些。然后我们将绘制地图 与这些相关的抗性基因和核RFLP标记。 转基因技术将被用来精确定位抗性基因。 序列分析将揭示其同一性。比如抗性基因 预计将包括这些药物的主要靶标的编码 提出问题，从而揭示他们的行动模式。这种方法是 适用于任何药物，并具有“无假设”的能力(否 需要提前对可能的目标进行猜测)并且速度很快。 到目前为止，我们有： 弓形虫物理连锁群的鉴定与鉴定 完成了核基因组的低分辨率遗传图谱。 绘制了与两种药物(新诺明和新诺明)耐药有关的基因 阿糖腺苷)，并开始生产第三种药物(阿托瓦酮)。 制定了弓形虫瞬时和稳定转染法。 识别特定药物的靶标对于了解如何 修改药物以提高其疗效，哪些其他药物可能有效，以及 哪些药物可能具有协同作用。

The overall goal of our work is to use modern genetics to identify the target(s) of new drugs that show promise for the treatment of toxoplasmosis in AIDS patients. Our approach has been to develop a genetic map and appropriate methodologies that will enable us to identify and clone genes that in mutant form can lead to resistance to a given drug. For the future, we plan to continue and extend this genetic approach to map resistance genes for atovaquone, clindamycin and other promising, new drugs as they become available. To do this, we will first expand our set of restriction-fragment-length-polymorphism (RFLP) probes to include those representing novel organella genomes. Then we will map the resistance genes with respect to these and the nuclear RFLP markers. Transfection will be used to precisely localize the resistance gene. Sequence analysis will reveal its identity. Such as resistance genes are expected to include those coding for the primary target of the drugs in question and thus reveal their mode of action. This approach is applicable to any drug and has the power of being "assumptionless" (no guesses need to be made in advance about the likely target) and rapid. To date, we have: identified and characterized the physical linkage groups of T. gondii completed a low resolution genetic map for the nuclear genome. mapped genes responsible for resistance to two drugs (sinefungin and adenosine arabinoside) and begun the process for a third (atovaquone). developed protocols for transient and stable transfection of T. gondii. Identifying the target of a given drug is essential to knowing how to modify the drug to improve its efficacy, what other drugs might work and which drugs might act synergistically.