谷氨酸棒状杆菌在乙酸盐和葡萄糖基质上的蓝白生长筛选(英文)

在谷氨酸棒状杆菌乙酸盐代谢调控关键酶-异柠檬酸裂解酶aceA基因两段启动子区域后面组装了无启动子的β-半乳糖苷酶报道基因,并且带上四环素抗性标记基因构建了两个检测质粒pROH7 和pROHO.通过该两个质粒转化了谷氨酸棒状杆菌野生型菌株Corynebacterium glutamicum ACTT 14752,分别获得了含有两个检测质粒的两种谷氨酸棒状杆菌,由此建立了谷氨酸棒状杆菌在乙酸盐和葡萄糖基质上的两个蓝白生长筛选系统.结果显示当两种菌株生长在含有葡萄糖的基本培养基上时呈现白色菌落,而生长在含有乙酸盐的基本培养基上时呈现蓝色菌落;其中菌落颜色还随菌株中检测质粒pROH7 和pROHO的不同而呈现淡蓝色和深蓝色的变化.含有检测质粒pROH7 和pROHO菌落的颜色变化反映了细胞在葡萄糖和乙酸盐不同和碳源上的调控状态,该两个质粒在鉴定乙酸盐代谢中的调控变化方面是有积极意义.

Blue-white colony screening on acetate and glucose in Corynebacterim glutamicum

To develop the blue-white colony screening with promoter test vector in Corynebacterium glutamicum,two vectors pROH7 and pROHO were constructed and transformed into Corynebacterium glutamicum ATCC 14752.They carry a tetracyclin resistance determinant and the promoterless β-galactosidase gene as a reporter gene behind two different promoter regions of the aceA gene of C.glutamicum,which encodes the enzyme isocitrate lyase,one of the key enzymes of acetate metabolism.Results were shown that white colonies grew on the minimum medium containing glucose,whearas blue colonies grew on the minimum medium containing acetate;The colour intensity varied from pale blue in colonies harbouring the test vector pROH7 to dark blue in colonies harbouring the test vector pROHO.The colour of the colonies with the test vectors pROH7 or pROHO reflects the regulatory state of the cells.Two vectors pROH7 and pROHO were shown to be functional in the identification of C.glutamicum clones with altered regulation of the acetate metabolism.``