机构地区: 广西大学生命科学与技术学院
出 处: 《广西农业生物科学》 2007年第B06期16-21,共6页
摘 要: 含有质粒pKD46的菌株BW25113,在L-阿拉伯糖诱导后,表达λ噬菌体的3个重组蛋白Exo、Bet和Gam,宿主菌就具有了同源重组的能力。利用λ噬菌体的Red重组系统构建了D-(+)-乳酸脱氢酶基因(ldhA)、乙醇脱氢酶基因(adhE)双突变的大肠杆菌BW25113C工程菌株和乙酸激酶基因(ackA)单突变的大肠杆菌BW25113工程菌株。将表达牛链球菌L-乳酸脱氢酶基因(ldhL)的pSE380质粒转化到BW25113C菌株中发酵培养35 h,用HLPC检测产物,尚未发现L-乳酸。 Plasmid pKD46 can express three proteins: Gam, get andExo. BW25113with pKD46 has the function of recombination when induced by L-arabinose. The two IdhA. adhE gene were deleted in Escherichia coli BW25113C and ackA was deleted in E. coli BW25113 using homologous recombination. The recombinant plasmid pSE380-Idh which expressed the L.-(+)-lactate dehydrogenase of Streptococcus bovis was transformed into E. coli BW25113C. The BW25113C/ pSE380-ldh was Shake-flask Ferment-cultivation for 35 h, using the HI.PC to detect the fermentation products, but had not fonnd the L-lactate yet.
领 域: [生物学]