CAJ | 학술논문

以受体菌的guaC基因为同源重组的指导序列,构建了整合表达载体pGT9GH,通过双交叉同源重组的方法将线性化的pGT9GH整合到受体菌的染色体上,构建了guaC基因的缺失突变株B.subtilis GJ07,并在guaC基因的5'端和3'端之间引入了1个拷贝核黄素操纵子,通过PCR方法验证了同源重组的正确性,最后测定了同源重组突变后的菌株GJ07的核黄素产量.结果表明:同源重组突变后的菌株GJ07的核黄素产量明显高于初始菌株GJ06,发酵60 h提高了24.5%.
이수체균적guaC기인위동원중조적지도서렬,구건료정합표체재체pGT9GH,통과쌍교차동원중조적방법장선성화적pGT9GH정합도수체균적염색체상,구건료guaC기인적결실돌변주B.subtilis GJ07,병재guaC기인적5'단화3'단지간인입료1개고패핵황소조종자,통과PCR방법험증료동원중조적정학성,최후측정료동원중조돌변후적균주GJ07적핵황소산량.결과표명:동원중조돌변후적균주GJ07적핵황소산량명현고우초시균주GJ06,발효60 h제고료24.5%.
An integrated expression vector pGT9GH was constructed by using guaC gene of receptor strain as guide sequence for homologous recombination. And then, by using the method of double crossover recombination between plasmid and chromosome, the linearized plasmid pGT9GH were integrated into B.subtilis GJ06 to get B.subtilis GJ07 with guaC gene mutant, in which, one copy of riboflavin operon was inserted between 5' and 3' end of guaC gene. The homologous recombination events were confirmed by PCR methods, and then, the riboflavin yield of mutant strain GJ07 was measured. The results of shake flask culture showed that the production of riboflavin of GJ07 was 24.5% higher than that of GJ06 in 60 h.