The Gram-positive bacterium Corynebacterium glutamicum represents a promising platform for the production of amino acids, organic acids, and other bio-products. However, the availability of only few expression vectors limits its use for production purposes, using metabolic engineering approaches when co-expression of several target genes is desired. To widen the scope for co-expression, the pCG1/p15A and pBL1/colE1 replicons were employed to construct the two differentially-inducible and compatible expression vectors pRG_Duet1 and pRG_Duet2. To functionally validate these newly constructed expression vectors, target genes for easily measurable enzymes were cloned and over-expression of these genes was investigated using respective enzyme assays. Furthermore, functionality and co-existence of the pCG1-based C. glutamicum - E. coli shuttle vector pRG_Duet1 were confirmed with pBL1-based expression vectors pRG_Duet2 and pEKEx2, using co-transformation and enzyme assays. The novel shuttle expression vectors pRG_Duet1 and pRG_Duet2 are attractive additions to the existing set of vectors for co-expression studies and metabolic engineering of C. glutamicum.

中文翻译：

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向前迈出了一步：在谷氨酸棒杆菌中基因共表达的兼容和双重诱导表达载体。

革兰氏阳性细菌谷氨酸棒杆菌代表了生产氨基酸，有机酸和其他生物产品的有前途的平台。但是，只有少数表达载体的可用性限制了其用于生产目的的使用，当需要多个靶基因的共表达时，使用代谢工程方法。为了扩大共表达的范围，使用了pCG1 / p15A和pBL1 / colE1复制子来构建两个差异诱导和兼容的表达载体pRG_Duet1和pRG_Duet2。为了在功能上验证这些新构建的表达载体，克隆了易于测量的酶的靶基因，并使用各自的酶测定法研究了这些基因的过表达。此外，基于pCG1的谷氨酸棒杆菌-E.的功能和共存。大肠杆菌穿梭载体pRG_Duet1已通过基于pBL1的表达载体pRG_Duet2和pEKEx2进行了共转化和酶法测定。新型穿梭表达载体pRG_Duet1和pRG_Duet2是现有谷氨酸棒杆菌共表达研究和代谢工程载体集合的引人注目的补充。