Michigan Biotechnology Institute and the Departments of Biochemistry and Microbiology, Michigan State University, East Lansing, Michigan 48824.
Appl Environ Microbiol. 1987 Jun;53(6):1387-90. doi: 10.1128/aem.53.6.1387-1390.1987.
To develop a biochemical genetic approach to understanding cell carbon synthesis or metabolic pathways in methanogens, Methanobacterium ivanovii was selected as a model organism for genetic manipulation studies. The organism displayed a colony size of 3 to 6 mm in less than 2 weeks and had a plating efficiency of about 90%, which made it suitable for replica plating. Mutagenesis and selection techniques were developed for selection of acetate auxotrophs. Chemical mutagenesis with ethyl methanesulfonate, followed by enrichment with bacitracin as a selective agent, resulted in stable acetate auxotrophs. M. ivanovii was very sensitive to UV, but UV-induced acetate auxotrophs were unstable and reverted within two to four transfers. The acetate auxotrophs were analyzed in relation to wild type for carbon monoxide dehydrogenase enzyme activity.
为了开发一种生化遗传学方法来理解产甲烷菌的细胞碳合成或代谢途径，选择 Methanobacterium ivanovii 作为遗传操作研究的模式生物。该生物在不到 2 周的时间内显示出 3 到 6 毫米的菌落大小，并且具有约 90%的平板效率，使其适合复制平板。为了选择乙酸营养缺陷型，开发了诱变和选择技术。用甲基磺酸乙酯进行化学诱变，然后用杆菌肽作为选择性试剂进行富集，导致稳定的乙酸营养缺陷型。M. ivanovii 对 UV 非常敏感，但 UV 诱导的乙酸营养缺陷型不稳定，在两到四次转移内回复。对野生型和一氧化碳脱氢酶酶活性相关的乙酸营养缺陷型进行了分析。
