Spindle pole body screen for fungicides
    9.
    发明授权
    Spindle pole body screen for fungicides 失效
    主轴杆体屏幕为杀菌剂

    公开(公告)号:US5418144A

    公开(公告)日:1995-05-23

    申请号:US66764

    申请日:1993-05-25

    IPC分类号: C12Q1/18 C12N1/00

    CPC分类号: C12Q1/18 Y10S435/942

    摘要: A method for the identification of agents which inhibit spindle pole body formation or function, thus exhibiting selective fungicidal activity, involves the incubation of test samples in cultures of a Saccharomyces cerevisiae strain that produces excess numbers of spindle pole bodies. Cultures containing samples that inhibit spindle pole bodies exhibit enhanced growth because the growing yeast is rescued from the adverse effects of excess spindle bodies. In the preferred practice of the invention, the test sample is added to a S. cerevisiae culture or culture area containing a strain that has a conditional mutation producing excess spindle pole bodies, such as diploid esp1-1 strains. The culture or culture area is preincubated under permissive conditions wherein the strain can grow to some extent, and then conditions are shifted to restrictive conditions so that the mutant strain either cannot grow or grows poorly. The extent of growth in the culture or culture area containing test sample is then compared with the extent of growth in the absence of test sample, and the presence of spindle pole body inhibition is determined by observation of whether culture growth in the presence of test sample exceeds growth in its absence. Preferred embodiments employ test samples on disks or in wells in solidified cultures, facilitating easy visual inspection of growth. A presumptive spindle pole body inhibitor, identified in random screening, is used as a positive control which can be compared to the test sample.

    摘要翻译: 用于鉴定抑制纺锤体体形成或功能,从而显示出选择性杀真菌活性的试剂的方法涉及在产生过量纺锤体的酿酒酵母菌株的培养物中孵育测试样品。 含有抑制锭子极体的样品的培养物表现出增强的生长,因为生长的酵母被从多余的纺锤体的不利影响中拯救出来。 在本发明的优选实践中,将测试样品加入到含有产生过量纺锤体的条件突变的菌株的酿酒酵母培养物或培养物区域中,例如二倍体esp1-1菌株。 培养或培养区域在允许条件下进行预培养,其中菌株可以在一定程度上生长,然后将条件转移到限制条件,使得突变菌株不能生长或生长差。 然后将含有测试样品的培养或培养区域的生长程度与不存在测试样品的生长程度进行比较,并通过观察测试样​​品存在培养生长来确定主轴极体抑制的存在 超过其缺席时的增长。 优选实施例在盘或在固化培养物中的孔中采用测试样品,便于生长的容易的视觉检查。 使用随机筛选鉴定的推定的主轴极体抑制剂作为阳性对照,可与测试样品进行比较。

    Screen for ultraspiracle inhibitors
    10.
    发明授权
    Screen for ultraspiracle inhibitors 失效
    超滤阻垢剂屏幕

    公开(公告)号:US6110698A

    公开(公告)日:2000-08-29

    申请号:US865960

    申请日:1997-05-30

    CPC分类号: C12Q1/02

    摘要: This invention discloses methods for identifying compounds, variant nuclear proteins and other auxiliary proteins that interfere with the Drosophila ultraspiracle protein ("Usp") and homologs thereof. The methods disclosed involve transformed yeast cells which contain a Usp binding partner, Usp or a homolog thereof which can bind with the Usp binding partner, and a reporter gene which requires a functional Usp--Usp binder partner complex for expression. The transformed yeast cells are incubated in the presence of a test compound to form a test culture and in the absence of a test compound to form a control culture. The expression of the reporter gene is monitored in one example by exposing the test and control cultures to canavanine under conditions in which control cultures exhibit reduced growth and detecting test cultures in which growth is increased relative to growth of control cultures.

    摘要翻译: 本发明公开了鉴定化合物,变体核蛋白和干扰果蝇超尖锐蛋白(“Usp”)及其同系物的其它辅助蛋白质的方法。 所公开的方法涉及含有可与Usp结合配偶体结合的Usp结合配偶体,Usp或其同系物的转化酵母细胞,以及需要功能性的Usp-Usp结合物配合物复合物进行表达的报告基因。 将转化的酵母细胞在测试化合物存在下孵育以形成测试培养物,并且在没有测试化合物的情况下形成对照培养物。 在一个实例中,通过将测试和对照培养物暴露于刀豆氨酸,在对照培养物显示减少的生长并检测其中生长相对于对照培养物的生长增加的测试培养物的条件下,监测报道基因的表达。