CAJ | 학술논문

目的:优化猪流行性腹泻抗原基因COE在乳酸乳球菌中的表达.方法:将表达猪流行性腹泻抗原基因COE的重组乳酸菌活化,酶切鉴定其稳定性,然后设计实验分别从pH值、温度、Nisin浓度、诱导时间、菌体密度等条件对COE表达进行优化,SDS-PAGE检测表达效果.结果:COE在乳酸乳球菌中的最佳表达条件为pH 7.0、T(温度)=30℃、Nisin=2ng/mL、t(诱导时间)=4h和OD_(600)=0.5,在以上条件下相对表达量分别达到了15.48%、15.05%、15.82%、14.72%和20.47%.在最佳表达条件下得到SOE的相对表达含量达到21%.结论:COE重组质粒稳定,其在乳酸菌中经优化表达后可为今后研制猪流行性腹泻乳酸菌疫苗提供数据.
목적:우화저류행성복사항원기인COE재유산유구균중적표체.방법:장표체저류행성복사항원기인COE적중조유산균활화,매절감정기은정성,연후설계실험분별종pH치、온도、Nisin농도、유도시간、균체밀도등조건대COE표체진행우화,SDS-PAGE검측표체효과.결과:COE재유산유구균중적최가표체조건위pH 7.0、T(온도)=30℃、Nisin=2ng/mL、t(유도시간)=4h화OD_(600)=0.5,재이상조건하상대표체량분별체도료15.48%、15.05%、15.82%、14.72%화20.47%.재최가표체조건하득도SOE적상대표체함량체도21%.결론:COE중조질립은정,기재유산균중경우화표체후가위금후연제저류행성복사유산균역묘제공수거.
Objective: To optimize expression of COE gene from porcine epidemic diarrhea virus(PEDV) in Lactococcus lactis.Method: The recombinant Lactococcus lactis that was inserted COE gene of PEDV was activated followed by identification its stability by digestion with endoenzyme.Then the COE gene was expressed and optimized from several aspects including different pH value of the medium,induction temperature(T),concentration of Nisin,induction time(t) and cell density.SDS-PAGE was processed to detect the expression.Result:The optimum expression condition results were shown for COE in Lactococcus lacti was pH 7.5,T 30℃,Nisin 2ng/mL,t 5h and OD_(600) 0.5,and the comparative expressing content under above condition reached to 15.48%,15.05%,15.82%,14.72% and 20.47%,respectively.Under the optimum expression condition the comparative expressing content of SOE reached to 21% in Lactococcus lactis.Conclusion: The recombinant COE plasmid was steadily existed in Lactococcus lactis and the optimization of expressing COE will supply new data to study engineering vaccine of PEDV later.