冷却牦牛肉贮藏过程中优势菌的 PCR-变性梯度凝胶电泳分析

应用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术,研究托盘保鲜膜包装的冷却牦牛肉在4℃贮藏过程中的微生物多样性及其动态变化.直接从样品中提取细菌总的DNA,采用降落PCR扩增16S rDNA的V3可变区序列,再通过DGGE得到动态变化的指纹图谱,并对主要条带进行测序分析.结果表明:检测到的优势腐败菌为Pseudomonas sp.(假单胞菌)、Lactococcus sp.(乳球菌)、Acinetobacter sp.(不动杆菌)、Brochothrix thermosphacta(热死环丝菌)、Enterobacteriaceae bacterium(肠杆菌科细菌),此外还检测到Uncultured Citrobacter sp.(非培养的柠檬酸杆菌)和Staphylococcus sp.(葡萄球菌)

PCR-DGGE Analysis of Dominant Bacteria in Yak Meat during Chilled Storage

In this study,polymerase chain reaction-denaturing gradient gel electrophoresis(PCR-DGGE) was used to investigate the changes in microbial diversity and dynamic changes of tray-packaged chilled yak meat during storage at 4 ℃.of Total bacterial DNA was directly extracted from meat samples.The touchdown PCR was applied to amplify the V3 region of the 16S rDNA gene before DGGE analysis.By analysis of the dynamic community directly obtained from the DGGE profiles and the DNA sequences of main bands,the predominant spoilage bacteria were found to be Pseudomonas sp.,Lactococcus sp.,Acinetobacter sp.,Brochothrix thermosphacta and Enterobacteriaceae bacterium.Uncultured Citrobacter sp.and Staphylococcus sp.were also identified in this study.