用于钝齿棒杆菌木糖代谢途径研究的基因工程菌构建

CONSTRUCTION OF GENETIC ENGINEERING STRAIN USED IN XYLOSE METABOLIC PATHWAY STUDY OF CORNEBACTERIUM CRENATUM

通过PCR方法从大肠杆菌基因组DNA中克隆得到木糖异构酶基因(xylA),将其连接到表达载体pXMJ19上,得到重组表达载体pXMJ19-xylA,通过电转化方法将pXMJ19-xylA转入宿主菌钝齿棒杆菌中,菌液PCR和重组质粒酶切结果证明,已成功构建了含大肠杆菌木糖异构酶基因的钝齿棒杆菌基因工程菌。该研究为钝齿棒杆菌利用木糖代谢途径生产琥珀酸及其代谢途径优化打下了基础。

Xylose isomerase gene （x-y/A） cloned from Escherichia coli genomic DNA by PCR was ligated to expression vector pXMJ19 to establish recombinant expression vectors pXMJ19-xy/A, which was transformed to host strain Corynebacterium crenatum by electrotransformation method. Colony PCR and recombinant plasmids digested results showed that C crenatum genetic engineering strain of harboring E.coli xylose isomerase gene has been constructed successfully. This study provides a foundation for producing succinic acid of C. crenatum xylose by sylose metabolic pathways and optimization of its metabolic pathway.