TETRAZOLIUM OVERLAY TECHNIQUE FOR POPULATION STUDIES OF RESPIRATION DEFICIENCY IN YEAST
TETRAZOLIUM OVERLAY TECHNIQUE FOR POPULATION STUDIES OF RESPIRATION DEFICIENCY IN YEAST
复制标题
DOI：
10.1126/science.125.3254.928
复制
发表时间：
1957-01-01
期刊：
影响因子：
56.9
通讯作者：
NAGAI, S
中科院分区：
文献类型：
--
作者：
OGUR, M;JOHN, RS;NAGAI, S
将四唑盐应用于酵母呼吸缺陷诊断的尝试是基于Raut的观察(1)，在四唑盐琼脂平板上发现的白色菌落是呼吸不足(Aer)，而红色菌落是呼吸充足(AER)。然而，Laskowski(2)证明了在2，3，5-三苯基四氮唑(TTC)存在的情况下生长诱导AER细胞产生Aer后代。最近，Nagai、Yanagishima、Hiraoka和Takada将TTC琼脂覆盖应用于在含有亚致死浓度铜的培养基上形成菌落后对铜敏感和抗铜的酵母细胞群的研究(3)。这项技术允许在短时间内使用高浓度的TTC，在此期间没有生长。这份报告描述了一种快速、直接诊断AER和Aer菌落的测试程序，在TTC琼脂上覆盖后。它的基本原理是使用相对较高浓度的TTC，在pH值足以在足够短的测试周期内在AER菌落中产生深红色，从而使AER菌落为白色。因此，可以在大多数菌株中进行诊断，而不需要考虑颜色模糊、生长抑制或指示剂诱导的Aer。试验用琼脂在0.067 M磷酸盐缓冲液中加入1.5%的BACTO琼脂，pH为7.0，TTC为0.1%。由于TTC通过与琼脂一起高压灭菌从化学上降低，所以单独的原料溶液被制备和高压灭菌。最终试验在550℃的无菌条件下混合，并储存在液体条件下的INCU中-所有技术论文和对它们的评论都发表在这一部分。稿件应以双倍行距打印，并提交一式两份。长度应限制在1200字以内；这包括插图或表格材料、参考文献和注释、作者(S)的姓名(S)和隶属关系(S)所占的空间。插图材料应限制在一个表格或一个数字内。所有解释性注释，包括对发表的感谢和授权，以及文献参考文献，应连续编号，输入正文，并放在文章末尾的“参考文献和注释”标题下。有关更详细的信息，请参阅1957年1月4日《科学》第125期，第16期(1957年1月4日)的《对投稿人的建议》。
Attempts to apply tetrazolium salts to the diagnosis of respiration deficiency in yeast are based on Raut's observation (1) that a white colony found on a tetrazolium agar plate was respiration deficient (aer), whereas red colonies were respiration sufficient (AER). Laskowski (2) demonstrated, however, that growth in the presence of 2, 3, 5-triphenyltetrazolium chloride (TTC) induced AER cells to produce aer offspring. Recently, Nagai, Yanagishima, Hiraoka, and Takada applied TTC agar overlays to the study of populations of copper-sensitive and copper-resistant yeast cells after colonies had formed on media containing sublethal concentrations of copper (3). This technique allowed the use of a high concentration of TTC in a short test period during which no growth occurred. This report describes a test procedure for the rapid, direct diagnosis of AER and aer colonies after overlay with TTC agar. Its rationale is based on the use of a relatively high concentration of TTC at a pH that yields deep red color de-velopment in AER colonies in a test period sufficiently short so that aer colonies are white. Diagnosis can thus be made in most strains without concern for color ambiguity, growth inhibition, or aer induction by the indicator. The test medium consists of 1.5 percent Bacto agar in 0.067 M phosphate buffer at pH 7.0, 0.1 percent with respect to TTC. Since theTTC is reduced chem-ically by autoclaving it with the agar, separate stock solutions are prepared and autoclaved. The final test agaris mixed under sterile conditions at 550C and is stored in the liquid condition in an incu-All technical papers and comments on them are published in this section. Manuscripts should be typed double-spaced and be submitted in duplicate. In length, they should be limited to the equivalent of 1200 words; this includes the space occupied by illustrative or tabular material, references and notes, and the author (s)'name (s) and affiliation (s). Illustrative materialshould be limited to one table or one figure. All explanatory notes, in-cluding acknowledgments and authorization for publication, and literature references are to be numbered consecutively, keyed into the text proper, and placed at the end of the article under the heading" References and Notes." For fuller details see" Suggestions to Contributors" in Science 125, 16 (4 Jan. 1957).