Possot O, Gernhardt P, Klein A, Sibold L
Département des Biotechnologies, Institut Pasteur, Paris, France.
Appl Environ Microbiol. 1988 Mar;54(3):734-40. doi: 10.1128/aem.54.3.734-740.1988.
The sensitivity of the methanogenic archaebacterium Methanococcus voltae to 12 inhibitors was tested in liquid medium. Four compounds appeared to be inhibitors of growth. Their MICs were as follows: pseudomonic acid, 0.1 micrograms/ml (0.19 microM); puromycin, 2 micrograms/ml (3.6 microM); methionine sulfoximine, 30 micrograms/ml (170 microM); and fusidic acid, 100 micrograms/ml (170 microM). On solid medium, the MICs were similar and the frequency of spontaneous resistance was found to be 5 X 10(-5) (methionine sulfoximine), 10(-7) (pseudomonic acid), and less than 10(-7) (puromycin and fusidic acid). Pseudomonic acid was found to inhibit isoleucyl-tRNA synthetase activity as measured by the in vitro aminoacylation of M. voltae tRNA with L-[U-14C]isoleucine. Fusidic acid and puromycin were shown to inhibit poly(U)-dependent polyphenylalanine synthesis in S30 extracts. Acetylpuromycin was inhibitory at much higher concentrations both in vivo and in vitro for M. voltae. Thus, the pac gene of Streptomyces alboniger, which is responsible for acetylation of puromycin and which conferred resistance to puromycin when introduced in eubacteria and eucaryotes, is a potential selective marker in gene transfer experiments with M. voltae. The latter was recently shown to be transformable. The same would be true for the cat gene of Tn9, which encodes resistance to fusidic acid in eubacteria in addition to resistance to chloramphenicol.
在液体培养基中测试了产甲烷古细菌沃氏甲烷球菌对12种抑制剂的敏感性。有四种化合物似乎是生长抑制剂。它们的最低抑菌浓度（MIC）如下：假单胞菌酸，0.1微克/毫升（0.19微摩尔）；嘌呤霉素，2微克/毫升（3.6微摩尔）；蛋氨酸亚砜亚胺，30微克/毫升（170微摩尔）；以及夫西地酸，100微克/毫升（170微摩尔）。在固体培养基上，MIC相似，且发现自发抗性频率为5×10⁻⁵（蛋氨酸亚砜亚胺）、10⁻⁷（假单胞菌酸），以及低于10⁻⁷（嘌呤霉素和夫西地酸）。发现假单胞菌酸可抑制异亮氨酰 - tRNA合成酶活性，这是通过用L - [U - ¹⁴C]异亮氨酸对沃氏甲烷球菌tRNA进行体外氨酰化来测定的。夫西地酸和嘌呤霉素被证明可抑制S30提取物中依赖于聚（U）的聚苯丙氨酸合成。乙酰嘌呤霉素在体内和体外对沃氏甲烷球菌均在高得多的浓度时具有抑制作用。因此，白色链霉菌的pac基因，该基因负责嘌呤霉素的乙酰化，并且当导入真细菌和真核生物中时赋予对嘌呤霉素的抗性，是在沃氏甲烷球菌的基因转移实验中的一个潜在选择标记。最近已证明后者是可转化的。Tn9的cat基因情况也是如此，该基因除了编码对氯霉素的抗性外，还编码真细菌中对夫西地酸的抗性。
