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利用嵌套 PCR-DGGE 技术分析日式和中式发酵豆酱中的细菌和真菌群落。

Analysis of bacterial and fungal communities in Japanese- and Chinese-fermented soybean pastes using nested PCR-DGGE.

机构信息

Kyung Hee University, Suwon, Republic of Korea .

出版信息

Curr Microbiol. 2010 May;60(5):315-20. doi: 10.1007/s00284-009-9542-4. Epub 2009 Nov 19.

Abstract

The microbial diversity of Japanese- and Chinese-fermented soybean pastes was investigated using nested PCR-denaturing gradient gel electrophoresis (DGGE). Five Japanese-fermented soybean paste samples and three Chinese-fermented soybean paste samples were analyzed for bacteria and fungi. Extracted DNA was used as a template for PCR to amplify 16S rRNA and 18S rRNA genes. The nearly complete 16S rRNA and 18S rRNA genes were amplified using universal primers, and the resulting products were subsequently used as a template in a nested PCR to obtain suitable fragments for DGGE. Tetragenococcus halophilus and Staphylococcus gallinarum were found to dominate the bacterial microbiota in Japanese samples, whereas Bacillus sp. was detected as the predominant species in Chinese samples. DGGE analysis of fungi in soybean pastes determined the presence of Aspergillus oryzae and Zygosaccharomyces rouxii in most of the Chinese and Japanese samples. Some differences were observed in the bacterial diversity of Japanese- and Chinese-fermented soybean pastes.

摘要

采用巢式 PCR-变性梯度凝胶电泳(DGGE)技术研究了日本和中国发酵大豆酱的微生物多样性。对 5 个日本发酵大豆酱样本和 3 个中国发酵大豆酱样本进行了细菌和真菌分析。提取的 DNA 用作 PCR 扩增 16S rRNA 和 18S rRNA 基因的模板。使用通用引物扩增了近乎完整的 16S rRNA 和 18S rRNA 基因,随后将所得产物用作巢式 PCR 的模板,以获得适合 DGGE 的片段。在日本样本中,发现四链球菌和鸡葡萄球菌主导细菌微生物群,而在中样品中检测到芽孢杆菌属为主要物种。通过对大豆酱中真菌的 DGGE 分析,确定了在大多数中国和日本样本中存在米曲霉和鲁氏酵母。在日本和中国发酵大豆酱的细菌多样性中观察到一些差异。

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