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目的通过对膳食诱导肥胖(Diet Induced Obesity,DIO)大鼠与健康对照组大鼠的肠道菌群结构的分析比较,寻找造成2组大鼠菌群结构差异的分子标识物,探讨肠道菌群的组成和结构与宿主肥胖之间的关系。方法ERIC-PCR结合Southern-blot得到肠道菌群基因组指纹图谱,利用Southern-blot与多元统计方法(PCA、PLS等)找出差异条带,回收差异条带进行测序,根据序列设计特异性引物,以定量PCR法对结果进行验证。结果ERIC-PCR图谱表明膳食诱导肥胖大鼠在肠道菌群结构上与正常大鼠存在着较大的区别;根据杂交结果找到一段基因组DNA片段为膳食诱导肥胖大鼠组所特有,定量分析表明该DNA片段在2组大鼠间区别明显。结论膳食诱导肥胖大鼠所特有的一段基因组DNA片段可作为肥胖大鼠肠道的特征分子标识物,该标识物有望用于膳食诱导肥胖的机制研究中。
OBJECTIVE: To analyze and compare the intestinal microflora in Diet Induced Obesity (DIO) rats and healthy control rats, and to find the molecular markers that cause the differences in the structure of the two groups of bacteria, The relationship between the composition and structure of the population and host obesity. Methods Genomic fingerprinting of intestinal flora was obtained by ERIC-PCR and Southern-blot. Southern blot and multivariate statistical methods (PCA, PLS, etc.) were used to identify the differential bands. The differential bands were recovered and sequenced. Based on the sequence specific primers , Quantitative PCR method to verify the results. Results The results of ERIC-PCR showed that there was a significant difference between the diet-induced obese rats and normal rats in the gut-induced obese rats. A genomic DNA fragment was found to be characteristic of the diet-induced obesity group according to the hybridization results. Quantitative analysis The DNA fragment was significantly different between the two groups of rats. Conclusion A segment of genomic DNA, which is characteristic of diet-induced obese rats, can be used as a marker of intestinal flora in obese rats. This marker is expected to be used in the mechanism of diet-induced obesity.