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題 名 | Molecular Discrimination of Lactobacillus plantarum Group Using Comparative Sequence Analysis of the dnaJ Gene and as a Target for Developing Novel Species-specific PCR Primers=利用dnaJ基因定序分析和種特異性PCR技術進行植物乳酸桿菌群相關菌種之分子鑑定 |
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作 者 | 黃建勳; 黃麗娜; 吳建平; 張慕慈; | 書刊名 | 中國畜牧學會會誌 |
卷 期 | 45:1 2016.03[民105.03] |
頁 次 | 頁45-55 |
分類號 | 368.87 |
關鍵詞 | 比較序列分析法; dnaJ基因; 植物乳酸桿菌群; 分子標的; 種特異性聚合酶連鎖反應; Comparative sequence analysis; dnaJ gene; Lactobacillus plantarum group; Molecular target; Species-specific two plex PCR; |
語 文 | 英文(English) |
中文摘要 | 本試驗旨在利用分生技術進行植物乳酸桿菌群( Lactobacillus plantarum group)之分子鑑別,包含以比較序列分析法分析 dnaJ基因,並開發種特異性引子供快速檢測。結果顯示,植物乳酸桿菌( Lb. plantarum)及其類緣關係相近菌種,皆可透過分析 dnaJ 基因序列清楚鑑別之,其種間序列相似度明顯低於 16S rRNA基因序列(84.6% vs. 99.0%),故 dnaJ基因序列可作為一個新穎分子標的,其可用於植物乳酸桿菌群之種層次鑑別。此外,基於 dnaJ 序列的高度變異性,設計兩組種特異性 PCR 引子,並搭配 two-plex 特異性 PCR 分析,可同時鑑定 Lb. plantarum與 Lb. pentosus。綜合得知,植物乳酸桿菌群相關菌種可明確被 dnaJ基因序列進行鑑別區分,又使用 two-plex 特異性 PCR 分析可快速、簡單和準確鑑定 Lb. plantarum與 Lb. pentosus。 |
英文摘要 | Identifying Lactobacillus plantarum and its closely related taxa at the species and subspecies level using only phenotypic and genotypic techniques often yields inaccurate results. In this study, the dnaJ gene was used for species discrimination by direct DNA sequencing and as target in a species-specific PCR assay. All examined strains were clearly distinguished from the closely related species by comparative sequence analysis of the dnaJ gene. The average sequence similarity for the dnaJ gene (84.6%) among type strains was significantly lower than that of the 16S rRNA sequence (99.0%) and was comparable to the current phylogenetic targets. Therefore, the dnaJ gene can be proposed as an additional molecular marker for species delineation of Lb. plantarum group that provides greater resolution than 16S rRNA. In addition, the species-specific primers were also developed based on the dnaJ gene sequences, which were then employed for two plex PCR using the template DNA of Lactobacillus strains. The PCR primer pairs were shown to be specific for Lb. plantarum and Lb. pentosus. Our data indicated that the phylogenetic relationships in the Lb. plantarum group were easily resolved by direct sequencing of the dnaJ gene, and the novel species-specific primers could be used to rapidly and accurately identify the species of Lb. plantarum and Lb. pentosus from the Lb. plantarum group using two plex PCR assays. |
本系統中英文摘要資訊取自各篇刊載內容。