α-淀粉酶基因在肠膜明串珠菌基因表达中的应用  

Study on application of α- amylase gene in the gene expression of Leuconostoc mesenteroides

作　　者：田云飞[1] 刘晓莉[1] 成文玉[1] 金红星[1] 

机构地区：[1]河北工业大学化工学院,天津300130

出　　处：《食品工业科技》2016年第10期203-207,共5页Science and Technology of Food Industry

摘　　要：目的:为了验证α-amy(α-淀粉酶基因)在肠膜明串珠菌中应用的可行性,构建重组质粒并建立了肠膜明串珠菌的三亲杂交体系。方法:以大肠杆菌-明串珠菌的穿梭载体p CW4为基础,构建了携带α-amy标记的重组质粒p CW7。肠膜明串珠菌ATCC8293(Leuconostoc mesenteroides)的g6ph(6-磷酸葡萄糖脱氢酶基因)表达盒插入到p CW7,并通过三亲杂交转化肠膜明串珠菌而获得重组菌株。结果:重组菌株的甘露醇产量比原始菌株提高了7%。三亲杂交中可以通过α-amy标记筛选重组子。结论:证实了α-amy标记在肠膜明串珠菌中应用的可行性。Objective： To verify the effect of the α- amy（ α- amylase gene） used in Leuconostoc mesenteroides,construction of recombined plasmid and triparental mating system in Leuconostoc mesenteroides were performed.Methods： The recombined plasmid p CW7,containing α- amy,was constructed based on E. coli- Leuconostoc shuttle vectors p CW4. 6- Phosphoglucose dehydrogenase gene（ g6ph） expression cassettes from Leuconostoc mesenteroides ATCC8293 was cloned into p CW7 and introduced into Leuconostoc mesenteroides by triparental mating. Results： Compared with the wild type,the recombination strain increased 7% yield of mannitol. The recombinant can be successfully selected by α- amylase gene markers.Conclusion： It showed the practicability ofα- amylase gene markers used in improving the gene expression of Leuconostoc mesenteroides.

关 键 词：肠膜明串珠菌 Α-淀粉酶基因 三亲杂交 6-磷酸葡萄糖脱氢酶 

分 类 号：TS201.3[轻工技术与工程—食品科学]