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 有希望治疗以下疾病的新药的靶点： 艾滋病患者的弓形虫病。 我们的方法是开发一个 基因图谱和适当的方法，使我们能够确定 并克隆基因，以突变的形式，可以导致抵抗给定的 药 在未来，我们计划继续并扩展这种基因 阿托伐醌、克林霉素和其他抗生素的耐药基因定位方法 有前途的新药。 为此，我们将首先 扩展我们的限制性片段长度多态性（RFLP）探针 包括那些代表新的细胞器基因组。 然后我们将绘制 与这些和核RFLP标记相关的抗性基因。 转染将用于精确定位抗性基因。 序列分析将揭示其身份。 如抗性基因， 预计将包括那些编码的主要目标的药物， 这是一个问题，并揭示了他们的行动方式。 这种方法 适用于任何药物，并有权力"无药可救"（没有 需要提前对可能的目标进行猜测）和快速。 迄今为止，我们已经： 鉴定并表征了T.弓形虫 完成了核基因组的低分辨率遗传图谱。 绘制了对两种药物（西芬净和 阿糖腺苷）并开始第三个（阿托伐醌）的过程。 开发了用于瞬时和稳定转染T.刚地。 确定给定药物的靶点对于了解如何 修改药物以提高其疗效，其他药物可能起作用， 哪些药物可能协同作用。

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.