Moschetti G, Villani F, Blaiotta G, Baldinelli A, Coppola S
Instituto di Microbiologia Agraria e Stazione di Microbiologia Industriale, Università degli Studi di Napoli Federico II, Portici, Italy.
FEMS Microbiol Lett. 1996 Nov 15;145(1):27-32. doi: 10.1111/j.1574-6968.1996.tb08552.x.
Sixty-four lactococcal strains isolated from natural whey starters were screened for the presence of the nisin structural gene by polymerase chain reaction. Seven of them showed a specific PCR product of 320 bp; only two produced antagonistic activity and were resistant to nisin. Southern blots of SmaI-digested DNA from PCR-positive strains hybridized with a nisA probe displayed a location of the gene on different SmaI fragments. Among PCR-positive strains, nisin producers showed specific transcript after reverse transcriptase-PCR, as well as some non-nisin-producing strains. The RT-PCR product could not be shown in one non-nisin-producing PCR-positive strain.
通过聚合酶链反应对从天然乳清发酵剂中分离出的64株乳球菌菌株进行了乳链菌肽结构基因存在情况的筛选。其中7株显示出320 bp的特异性PCR产物;只有2株产生拮抗活性且对乳链菌肽具有抗性。用nisA探针与PCR阳性菌株经SmaI酶切的DNA进行Southern杂交,结果显示该基因位于不同的SmaI片段上。在PCR阳性菌株中,乳链菌肽产生菌在逆转录PCR后显示出特异性转录本,一些非乳链菌肽产生菌也如此。在一株非乳链菌肽产生的PCR阳性菌株中未检测到RT-PCR产物。