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利用PCR-DGGE技术分析桑天牛成虫肠道菌群结构及优势菌群,获取桑天牛肠道微生物的多样性信息。从桑天牛成虫肠道中提取细菌基因组DNA,以细菌16S rDNA基因通用引物27F/1495R和27F/519r+GC进行V3可变区PCR扩增,将长约500 bp的扩增产物经变性梯度凝胶电泳(DGGE)分离后,进行优势条带分析、DNA回收、克隆、测序等,初步得到分别属于肠杆菌属(Enterobacter)、不动杆菌属(Acinetobacter)、埃希氏菌属(Escherichia)、志贺菌属(Citrobacter)、克雷伯氏菌属(Klebsiella)、柠檬酸菌属(Shigella)、泛菌属(Pantoea)和沙雷氏菌属(Serratia)的8个细菌菌株,其中优势细菌为克雷伯氏菌属的细菌菌株,其次是沙雷氏菌属的细菌菌株。将获取的细菌16S rDNA序列在GenBank数据库中进行BLAST比对分析,相似度在97%以上的有6个菌株,其中有5个菌株与传统方法分离菌株的鉴定结果一致,表明基于16S rDNA的PCR-DGGE技术可用于桑天牛肠道菌群多样性研究。
Using PCR-DGGE technology to analyze the intestinal flora of Morinda spp. Adults and its dominant flora, and gain the diversity information of intestinal microorganisms of Morinda. Bacterial genomic DNA was extracted from the intestinal tract of adult Morus alba, followed by PCR amplification of the V3 variable region of bacterial 16S rDNA gene universal primers 27F / 1495R and 27F / 519r + GC. The amplified product, about 500 bp in length, After separation by gel electrophoresis (DGGE), the dominant bands, DNA recovery, cloning, sequencing and so on were obtained, which were classified into Enterobacter, Acinetobacter, Escherichia, 8 bacterial strains of Citrobacter, Klebsiella, Shigella, Pantoea and Serratia, among which the predominant bacteria are Bacterial strains of Klebsiella, followed by bacterial strains of Serratia. The obtained bacterial 16S rDNA sequences were analyzed by BLAST in the GenBank database, 6 strains with similarity above 97%, of which 5 strains were consistent with those of the traditional method, indicating that 16S rDNA based PCR DGEGE Technique Can Be Used to Study Diversity of Intestinal Microflora in Morus alba.