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在质粒pUC19中插入菊欧氏杆菌pehX启动子,并与全合成的运动发酵单胞菌的丙酮酸脱羧酶基因(pdc),乙醇脱氢酶基因(adhB)串联,构建pUC19-pdc-adhB乙醇发酵重组克隆,转化大肠杆菌DH5α,经气相色谱检测,在大肠杆菌中乙醇的表达量达到0.85%。菊欧氏杆菌富含多种纤维素酶,与一些仅可利用基本糖类作为碳源的大肠杆菌等相比,其可以利用纤维素作为碳源。本实验设计将pUC19-pdc-adhB乙醇发酵重组克隆转化至菊欧氏杆菌,尝试用氯化钙、电击等转化方法,但均没有获得转化子。提示需要寻找新的转化方法或者将pdc和adhB基因整合到菊欧氏杆菌的基因组中以使其能产生乙醇。
The plasmid pUC19 was inserted into the Ehrlichia pehX promoter and ligated with the fully synthesized Z. mobilis pyruvate decarboxylase gene (pdc) and the alcohol dehydrogenase gene (adhB) to construct pUC19-pdc-adhB ethanol The recombinant plasmid was transformed into Escherichia coli DH5α and detected by gas chromatography. The expression of ethanol in E. coli reached 0.85%. Ehrlichia is rich in a variety of cellulases, which can utilize cellulose as a carbon source compared to some Escherichia coli and the like that can utilize only basic sugars as a carbon source. In this experiment, we transformed the recombinant pUC19-pdc-adhB ethanol into E.coli and tried to use calcium chloride, electric shock and other transformation methods, but no transformants were obtained. Suggesting the need to find new transformation methods or the integration of the pdc and adhB genes into the genome of Elymus so that it can produce ethanol.