Corynebacterium glutamicum is an important industrial strain used for the production of amino acids and vitamins. Most tools developed for overexpression of genes in C. glutamicum are based on the inducible promoter regulated by the lacIq gene or contain an antibiotic resistance gene as a selection marker. These vectors are essential for rapid identification of recombinant strains and detailed study of gene functions, but, as a considerable disadvantage, these vectors are not suitable for large-scale industrial production due to the need for the addition of isopropyl-β-D-thiogalactopyranoside (IPTG) and antibiotics. In this study, the novel Escherichia coli-C. glutamicum shuttle expression vector pLY-4, derived from the expression vector pXMJ19, was constructed. The constitutive vector pLY-4 contains a large multiple cloning site, the strong promoter tacM and two selective markers: the original chloramphenicol resistance gene cat is used for molecular cloning operations, and the auxotrophy complementation marker alr, which can be stably replicated in the auxotrophic host strain without antibiotic selection pressure, is used for industrial fermentation. Heterologous expression of the gapC gene using the vector pLY-4 in C. glutamicum for L-methionine production indicated the potential application of pLY-4 in the development of C. glutamicum strain engineering and industrial fermentation.

中文翻译：

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具有营养缺陷型互补系统的谷氨酸棒杆菌的新型表达载体。

谷氨酸棒杆菌是用于生产氨基酸和维生素的重要工业菌株。为在谷氨酸棒杆菌中基因过度表达而开发的大多数工具都基于由lacIq基因调节的诱导型启动子，或包含抗生素抗性基因作为选择标记。这些载体对于快速鉴定重组菌株和详细研究基因功能是必不可少的，但是，作为一个重大缺点，由于需要添加异丙基-β-D-硫代吡喃半乳糖苷，这些载体不适合大规模工业生产（IPTG）和抗生素。在这项研究中，新型大肠杆菌-C。构建了来自表达载体pXMJ19的谷氨酸穿梭表达载体pLY-4。组成型载体pLY-4包含一个大型的多克隆位点，强启动子tacM和两个选择标记：最初的氯霉素抗性基因cat用于分子克隆操作，营养缺陷型互补标记alr可在营养缺陷型宿主菌株中稳定复制而无抗生素选择压力，用于工业发酵。使用载体pLY-4在谷氨酸棒杆菌中异源表达gapC基因以生产L-甲硫氨酸表明pLY-4在谷氨酸棒杆菌菌株工程开发和工业发酵中的潜在应用。用于工业发酵。使用载体pLY-4在谷氨酸棒杆菌中异源表达gapC基因以生产L-甲硫氨酸表明pLY-4在谷氨酸棒杆菌菌株工程开发和工业发酵中的潜在应用。用于工业发酵。使用载体pLY-4在谷氨酸棒杆菌中异源表达gapC基因以生产L-甲硫氨酸表明pLY-4在谷氨酸棒杆菌菌株工程开发和工业发酵中的潜在应用。