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首页> 外文期刊>Journal of medical bacteriology. >A New Approach for Designing a Potentially Vaccine Candidate against Urinary Tract Infection by Using Protein Display on Lactobacillus
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A New Approach for Designing a Potentially Vaccine Candidate against Urinary Tract Infection by Using Protein Display on Lactobacillus

机译：通过使用乳酸菌上的蛋白质展示设计针对尿路感染的潜在疫苗候选者的新方法

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Background: The prevalence of Urinary Tract Infection (UTI) is really high in the world. Escherichia coli is a major agent of UTI. One of the strategies for decreasing UTI infections is vaccine development. As the attachment is a really important stage in colonization and infection, at- tachment inhibition has an applied strategy. FimH protein is a major factor during bacterial colonization in urinary tract and could be used as a vaccine. Thus, it was considered in this research as a candidate anti- gen.Methods: The sequences of fimH and acmA genes were used for de- signing a synthetic gene. It was cloned to pET23a expression vector and transformed to E. coli (DE3) Origami. To confirm the expression of recombinant protein, SDS-PAGE and western blotting methods were used. Subsequently, recombinant protein was purified. On the other hand, Lactobacillus reuteri was cultured and mixed with FimH / AcmA recombinant protein. The rate of protein localization on lactobacillus surface was assessed using ELISA method.Results: It was showed that the recombinant protein was expressed in E. coli (DE3) Origami and purified by affinity chromatography. More- over, this protein could be localized on lactobacillus surface by 5 days. Conclusion: In current study, a fusion recombinant protein was pre- pared and displayed on L. reuteri surface. This strain could be used for animal experiment as a competitor against Uropathogenic E. coli (UPEC). Using manipulated probiotics strains instead of antibiotic ther- apy could decrease the antibiotic consumption and reduce multi-drug resistant strains.
机译：背景：世界上泌尿道感染（UTI）的患病率确实很高。大肠杆菌是尿路感染的主要药物。减少UTI感染的策略之一是疫苗开发。由于依恋是定殖和感染的真正重要阶段，因此抑制依恋已成为一种应用策略。 FimH蛋白是细菌在尿道中定植的主要因素，可以用作疫苗。因此，在这项研究中它被认为是候选抗原。方法：使用fimH和acmA基因的序列设计合成基因。将其克隆至pET23a表达载体，并转化至大肠杆菌（DE3）折纸。为了证实重组蛋白的表达，使用了SDS-PAGE和western blotting方法。随后，纯化重组蛋白。另一方面，培养罗伊氏乳杆菌并与FimH / AcmA重组蛋白混合。结果：表明重组蛋白在大肠杆菌（DE3）Origami中表达，并经亲和层析纯化。而且，这种蛋白质可以在5天后定位在乳酸菌表面。结论：目前的研究中，制备了一种融合重组蛋白并在罗伊氏乳杆菌表面上展示。该菌株可作为动物实验对抗尿毒症性大肠杆菌（UPEC）。使用经操作的益生菌菌株代替抗生素治疗可减少抗生素的消耗并减少多药耐药菌株。

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