耐辐射球菌抑菌圈试验过程中出现的问题与分析

Analysis on Problems Appeared in Bacteriostatic Ring Experiment of Deinococcus radiodurans

[目的]分析耐辐射球菌抑菌圈试验过程中出现的问题并提出解决方法。[方法]采用已发表的基因组序列作参考,对来自于耐辐射球菌的所有菌株均用TGY培养基或者TGY平板在30℃条件下进行培养,参照Debabrota等的方法进行抑菌圈试验,所用突变体源于以前的试验。[结果]分析认为,在做耐辐射球菌抑菌圈试验的过程中,首先需要注意的是严格灭菌,每次重新拿到超净工作台上的离心管和钢圈都要进行严格的灭菌处理。试验前对上清液要进行高速离心,以防止在上清液中混有菌体,从而造成结果不准确。最好在进行抑菌圈试验以前,先对上清液中是否含有菌体做一个前期试验,以免造成后面不必要的麻烦。[结论]该研究所提出的解决方法对做好以后的试验具有借鉴意义。

[ Objective ] The study aimed to analyze the problems apDeared in bacteriostatie ring experiment of Deinoeoccus radiodurans and propose the resolving method. [ Method] With the reported genome sequence as the reference, the all strains from D. radiodurans were cultured on the TGY medium or on the TGY plate at 30℃ and the bacteriostatic ring experiment was conducted referring to the method made by Debabrota et at, with the mutant being originated from the fimner test. [ Result ] On base of the analysis it was suggested that during the bacteriostatic ring experiments of D. radiodurans, sterilizing strictly was the most important at first and the sterilizing must be. performed before these microtubes and iron rings were denovo brought into the bench every time. The superuatant should be made for high speed centrifugation before experiment, which can prevent the supernatant having any bacteria and ensure the accurate testing result. Before the supernatant was used for bacteriostatic ring experiment, it was better to make the prophase test to confirm that if the supernatant has the bacteria so as to ensure no bacteria was contained in the supernatant prior to formal experiment. [ Conclusion]The suggestion proposed by this study had reference meaning for doing well the latter ecperiment.