CAJ | 학술논문

质粒消除是签定质粒和获得无质粒菌株的重要方法,是乳酸菌进行遗传学改造所需的一项重要技术.试验采用高温和消除剂结合的方法,对乳酸乳球菌镉抗性菌株进行质粒消除,探讨温度、消除剂吖啶橙的用量和作用时间对乳酸乳球菌镉抗性质粒消除的影响.结果表明,39℃高温可以质粒消除,而37和41℃均无此效果;独自吖叮橙作用未获得质粒消除菌株;39℃高温-吖啶橙同时作用比高温-吖啶橙交替作用消除率高,而39℃高温-20 μg·mL-1吖啶橙共同作用12 d,消除率可达98%.根据消除结果,以疑似功能性质粒为模板,进行PCR扩增,获得预期片段,进一步证实了其功能.
질립소제시첨정질립화획득무질립균주적중요방법,시유산균진행유전학개조소수적일항중요기술.시험채용고온화소제제결합적방법,대유산유구균력항성균주진행질립소제,탐토온도、소제제아정등적용량화작용시간대유산유구균력항성질립소제적영향.결과표명,39℃고온가이질립소제,이37화41℃균무차효과;독자아정등작용미획득질립소제균주;39℃고온-아정등동시작용비고온-아정등교체작용소제솔고,이39℃고온-20 μg·mL-1아정등공동작용12 d,소제솔가체98%.근거소제결과,이의사공능성질립위모판,진행PCR확증,획득예기편단,진일보증실료기공능.
Plasmid elimination is a key method of plasmid identificaton and obtaining non-plasmid strain, and it is an important technology which was needed by genetics transformation in lactic acid bacteria. High temperature and the elimination agent were combined by plasmid elimination for a cadmium-resistant Lactococcus lactis strains, the amount and the time by using temperature and eUminaton agent acridine orange (AO) were approached on the effect of cadmium-resistant Lactococcus lactis plasmid. The results showed that 39℃ high temperature could eliminate the plasmid, but 37 and 41℃ had no effects. Plasmid elimination strain was not obtained by using AO alone, the elimination rate by the method of both high temperature and AO at the same time was higher than that of by interchanging between high temperature and AO. The combined effect of 39℃ temperature and 20 μg·mL-1 AO for 12 days was significant, the elimination rate was as high as 98%. According to the results, PCR amplification was based on the suspected functional plasmid as a template, achieved the desired fragment, and further confirmed its functions.