摘要
比较研究冷却猪肉不同前处理对细菌DNA提取及PCR-变性梯度凝胶电泳(DGGE)的影响。冷却猪肉4℃贮藏至第5天时,分别采用冻融、超声、摇床和匀浆前处理后,提取细菌DNA,进行Dilution-PCR和DGGE分析。结果表明,不同前处理方法所制备的DNA量稍有差别,但对PCR-DGGE结果无显著影响,故可根据实际情况选择上述不同前处理方法。同时对16S rDNA V3区的DGGE图谱进行割胶测序,检测到腐败菌主要是假单胞菌属,其他还有不动杆菌属、环丝菌属和沙雷氏菌属。
Dilution-PCR and PCR-DGGE were used to evaluate effect of four pre-treatment methods such as freeze-thaw,ultrasonification,shake and homogenization on extraction of bacterial DNA from chilled pork in this study.After 4 days of storage at 4 ℃,chilled pork was subjected to pre-treatment by the above methods for bacterial DNA extraction followed by dilution-PCR and PCR-DGGE analyses.Results showed that four pretreatment methods results in a slight difference in bacterial DAN yield but had no notable effect on PCR-DGGE analysis.Therefore,these pretreatment methods could be chosen according to actual requirement.Sequence analysis of DGGE bands of 16S rRNA gene V3 region showed that the predominant spoilage bacterium in chilled pork was Pseudomonas and others including Acinetobacter,Brochothrix,and Serratia were also detected.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2010年第6期258-262,共5页
Food Science
基金
国家"863"计划重点项目(2008AA100804)
江苏省高校自然科学基础研究项目(08KJB550005)
上海市科委重大攻关项目(07dz19508)
