摘要
目的应用PCR和DGGE技术分析饲喂鲎素对小鼠粪样细菌区系变化的影响。方法将健康21日龄清洁级KM小鼠,随机分为4个处理,分别饲喂生理盐水、合成鲎素、抗生素和益生菌。粪样细菌16SrDNA的V6-V8可变区经PCR扩增,扩增产物经DGGE电泳后再进行相似性分析。切下9条DGGE胶中特异性条带,PCR扩增,TA克隆,测序,鉴定细菌种属。结果对照组DGGE电泳条带较多,在不同时期肠道菌群相似性均较高,相似性在73.5%～76.7%;灌胃合成鲎素和益生菌悬液组,DGGE电泳条带数和对照组比略有减少,不同处理时期肠道菌群相似性在68.1%～69.4%;灌胃抗生素组,DGGE电泳条带数明显减少,不同处理时期肠道菌群相似性在51.4%～63.0%。结论断奶小鼠在灌胃鲎素和抗生素后,肠道微生物区系和对照比发生了改变,最终形成特定的微生物区系;DGGE中特异条带的鉴定表明,灌胃鲎素可以促进粪链球菌和乳酸杆菌的生长,提高数量。
Objective To monitor the development of bacterial community in faeces of rats administered intragastrically with peptides by PCR and denaturing gradiem gel electrophoresis {DGGE）. Method KM rats were randomly assigned to 4 groups of 12 mice in each, and each group was administered for 21d intragastrically with physiological saline, synthetical tachyplesin, antibiotics and probiotic respectively, 0.5 ml of administration for each rat. The regions V6 to V8 of the 16S rDNA of faecal bacteria were amplified. DGGE profiles of the PCR amplicons were compared by similarity analysis. Nine specific DGGE bands were excised and the DNA associated with each of these bands was recovered and amplified by PCR. PCR amplieons were then cloned into the Vector pUCM-T. The inserts were sequenced. The bacterial taxa represented by the DGGE bands the were determined. Result The DGGE bande of the rats administered with taehyplesin and ehloramphenieol were decreased obviously; DGGE similarity were at 68.1% ~ 69.4% and 51.4% ~ 63.0% respeetlvely. Conclusion The growth of Streptococcus faecium can be promoted by treatment with taehyplesin. It suggests that taehyplesin are capable of modifying the bacterial composition of the caeeal content in rats.
出处
《中国微生态学杂志》
CAS
CSCD
2010年第11期982-987,共6页
Chinese Journal of Microecology
基金
安徽省高校自然科学研究资助项目(KJ2009B014)
安徽省高等学校省级食品科学与工程特色专业建设点资助(20101091)
安徽省食品科学与工程教学团队项目资助(20101094)
蚌埠学院优秀人才基金项目
关键词
鲎素
DGGE
肠道
菌群
Tachyplesin
DGGE
Intestinal tract
Bacterial community
