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目的利用不同的内切酶和电泳条件评价非脱羧勒克菌的脉冲场凝胶电泳(PFGE)实验方法,建立一系列非脱羧勒克菌的PFGE方法。方法 2013—2015年从甘肃省某婴儿奶粉加工企业车间的环境和原料中分离非脱羧勒克菌,利用溶菌酶和蛋白酶K对其裂解后,选用不同的内切酶对胶块内的核酸进行酶切,脉冲场电泳后评价分析电泳图谱,得到最佳实验方案。结果经过几种内切酶的对比试验后,发现SpeⅠ酶在本试验方案中反应稳定,酶切后条带大约为17~21条,且分布均匀,更适合用于图谱分析。结论本研究确定了非脱羧勒克菌的PFGE方法,可以为非脱羧勒克菌在食品、药品等加工过程的污染以及医院感染监测,提供溯源技术手段。
OBJECTIVE: To evaluate the PFGE method of non-decarboxylated lecce bacteria using different endonuclease and electrophoresis conditions. Methods From 2013 to 2015, non-decarboxylated Lactobacillus from decoction of lysozyme and proteinase K was isolated from the environment and raw materials of a baby milk powder processing enterprise workshop in Gansu Province. After cleavage with lysozyme and proteinase K, different endonucleases were used for the nucleic acid in the gel block Enzyme digestion, pulsed field electrophoresis after electrophoresis analysis, the best experimental program. Results After several comparison tests of endonucleases, SpeⅠase was found to be stable in this experimental protocol. The bands after digestion were about 17-21, and were evenly distributed, which was more suitable for the analysis of the spectrum. Conclusion This study determined the PFGE method of non-decarboxylated bacterium, which could provide the traceability technology for the non-decarboxylated bacteriocin in the process of food, medicine and other pollution and hospital infection monitoring.