鼠李糖乳杆菌Probio-M9的特异性荧光原位杂交探针设计及应用
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国家自然科学基金项目(31720103911);内蒙古科技重大专项(ZDZX2018018)


Design and Application of a Specific Fluorescent in Situ Hybridization Probe for Lactobacillus rhamnosus Probio-M9
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    摘要:

    宿主口服益生菌的定性和局部检测具有重要意义。本研究通过设计株水平特异性荧光原位杂交(FISH)探针来研究鼠李糖乳杆菌在宿主肠道中的定殖。根据鼠李糖乳杆菌Probio-M9和其它相关物种的基因组序列,分别设计了鼠李糖乳杆菌种水平特异性探针(Probe-16S)和鼠李糖杆菌Probio-M9菌株水平特异性探针(Probe-SP)。在使用15株鼠李糖乳杆菌菌株和15株其它益生菌验证这些特异性探针后,通过荧光原位杂交(FISH)验证鼠李糖乳杆菌Probio-M9菌株的特异性探针在体外和体内的适用性。结合荧光D-氨基酸(FDAA)探针,在大鼠的肠道中检测到鼠李糖乳杆菌Probio-M9的活细胞。本研究设计的FISH探针可从肠道菌群中特异性鉴定鼠李糖乳杆菌Probio-M9,该方法可与FDAA探针结合使用,以进一步探索该益生菌菌株在宿主肠道中的定位。

    Abstract:

    Qualitative and localized detection of oral probiotics in the host has great significance. In this study, a horizontal specific fluorescence in situ hybridization probe was designed to study the colonization of Lactobacillus rhamnosus in the host intestinal tract. Based on the genome sequences of L.rhamnosus Probio-M9 and other related species, the L. rhamnosus species-level specific probe (Probe-16S) and the L.rhamnosus Probio-M9 strain-level specific probe (Probe-SP) were designed. After these specific probes verified using 15 strains of Lactobacillus rhamnosus and 15 other probiotics of the genus, the L.rhamnosus Probio-M9 strain-level specific probe was examined its applicability both in vitro and in vivo by fluorescence in situ hybridization (FISH). Finally, combining a fluorescent D-amino acid (FDAA)-based metabolic probe, the living cells of L.rhamnosus Probio-M9 were detected in the intestinal tract of rats. The strain-level FISH probe we designed could specifically identify the L.rhamnosus Probio-M9 from the gut microbiota. This method could combine with the FDAA probes to provide a reliable technic for further exploring the translocation of this probiotic strain in the host.

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徐瑞波;高广琦;白晓晔;李伟程;张和平.鼠李糖乳杆菌Probio-M9的特异性荧光原位杂交探针设计及应用[J].中国食品学报,2021,21(11):230-235

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  • 在线发布日期: 2021-12-17
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