Schleyer Ute, Bringer-Meyer Stephanie, Sahm Hermann
Institut für Biotechnologie 1, Forschungszentrum Jülich GmbH, D-52425 Jülich, Germany.
Int J Food Microbiol. 2008 Jun 30;125(1):91-5. doi: 10.1016/j.ijfoodmicro.2007.04.016. Epub 2007 Sep 4.
Gluconobacter oxydans is known for causing rapid and incomplete oxidation of a wide range of sugars, sugar acids and sugar alcohols. Therefore, this microorganism is already employed in several biotechnological processes that involve incomplete oxidation of a substrate, e.g. vitamin C or dihydroxyacetone production. To fully exploit the oxidative potential of G. oxydans, characterization of the biological role of gene products is essential. To take advantage of the genome sequence of G. oxydans DSM 2343, based on pBBR1MCS5, we constructed a new cloning and expression vector. The newly established vector pEXGOX will significantly decrease duration of cloning and increase cloning efficiency. It has the following advantages: (i) small size (5.7 kbp); (ii) complete sequence; (iii) variety of unique restriction sites; (iv) direct cloning of PCR products; (v) strong promoter. The pEXGOX plasmid was successfully used to clone G. oxydans genes and has the potential to facilitate studies of gene function of several G. oxydans open reading frames.
氧化葡萄糖杆菌以能快速且不完全地氧化多种糖类、糖酸和糖醇而闻名。因此，这种微生物已被应用于多种涉及底物不完全氧化的生物技术过程，例如维生素C或二羟基丙酮的生产。为了充分发掘氧化葡萄糖杆菌的氧化潜力，对基因产物的生物学作用进行表征至关重要。为了利用氧化葡萄糖杆菌DSM 2343的基因组序列，基于pBBR1MCS5，我们构建了一种新的克隆和表达载体。新建立的载体pEXGOX将显著缩短克隆时间并提高克隆效率。它具有以下优点：（i）尺寸小（5.7 kbp）；（ii）序列完整；（iii）有多种独特的限制性酶切位点；（iv）可直接克隆PCR产物；（v）启动子强。pEXGOX质粒已成功用于克隆氧化葡萄糖杆菌基因，并且有潜力促进对氧化葡萄糖杆菌多个开放阅读框的基因功能研究。
