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沙门氏菌是一类常见的肠道致病菌，广泛存在于自然界中，尤其是动物体内，是食品安全和公共卫生领域中的重要病原。鼠伤寒沙门氏菌（Salmonella enterica serovar Typhimurium）是沙门氏菌的一个典型亚种，能够感染多种动物，包括小鼠和人类。抗生素治疗虽然可以有效抑制细菌生长并缓解症状，但使用不当会导致耐药性问题甚至造成肠道微生态失衡，必须谨慎使用并探索替代治疗方法，如益生菌治疗。鉴于此，本研究以干酪乳杆菌（Lactobacillus casei ATCC393）为宿主益生菌，将ST6GALNAC1、LFCA和TFF3基因融合，定向克隆至pPG-612载体，经电转化得到新功能型干酪乳杆菌Lc-pPG-ST6-LFCA-TFF3（Lc-pPG-SLT）。通过细胞安全性实验鉴定工程菌对肠上皮细胞LS174T的安全性，通过实时荧光定量PCR（RT-qPCR）检测，发现工程菌能显著上调紧密连接蛋白Occludin和Claudin-1的表达，其自身并未诱导渗漏型孔道蛋白Claudin-2的异常升高，同时能够抑制炎症因子的表达。该菌株的构建为开发靶向肠道屏障修复的基因工程益生菌提供了实验依据和新思路。
Abstract：Salmonella is a common class of intestinal pathogenic bacteria widely present in nature， particularly within animal hosts， and represents a significant pathogen in the fields of food safety and public health. Salmonella enterica serovar Typhimurium (S. Typhimurium) is a typical serovar capable of infecting a variety of animals， including mice and humans. Although antibiotic treatment can effectively inhibit bacterial growth and alleviate symptoms， improper use may lead to drug resistance and even disrupt intestinal microecological balance. Therefore， antibiotics must be used with caution， and alternative therapeutic approaches， such as probiotic therapy， should be explored. In view of this， the present study utilized Lactobacillus casei ATCC 393 as the host probiotic strain. The ST6GALNAC1， LFCA， and TFF3 genes were fused and directionally cloned into the pPG-612 vector. The resulting recombinant plasmid was introduced into L. casei via electroporation to generate a novel functional probiotic strain， designated as Lc-pPG-ST6-LFCA-TFF3 (Lc-pPG-SLT). Cellular safety assays were conducted to evaluate the biosafety of the engineered strain towards intestinal epithelial LS174T cells.RT-qPCR analysis revealed that the engineered strain significantly upregulated the expression of tightening junction proteins Occludin and Claudin-1， while it did not induce the abnormal elevation of the leaky protein Claudin-2 typically caused by pathogenic invasion， and simultaneously suppressed the expression of inflammatory factors. The development of engineered strain provides an experimental basis and a novel strategy for the development of genetically engineered probiotics targeting intestinal barrier repair.
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基本信息:
中图分类号:TS201.3;S852.6
引用信息:
[1]邵鑫,张璇,单裕荃,等.黏膜修复型干酪乳杆菌Lc-ST6-LFCA-TFF3的构建及特性研究[J].经济动物学报().
基金信息:
吉林省科技发展计划项目（20220101311JC）
2026-05-25
2026-05-25
2026-05-25