A food-grade fimbrial adhesin FaeG expression system in Lactococcus lactis and Lactobacillus casei
A food-grade fimbrial adhesin FaeG expression system in Lactococcus lactis and Lactobacillus casei
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DOI：
10.1139/cjm-2015-0596
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发表时间：
2016-03-01
影响因子：
2.8
通讯作者：
Kong, J.
中科院分区：
文献类型：
--
作者：
Lu, W. W.;Wang, T.;Kong, J.
产肠毒素大肠杆菌（ETEC）感染是新生仔猪腹泻的主要原因。菌毛作为 ETEC 发病机制中的定植因子，构成了 ETEC 疫苗接种的主要目标。乳酸菌（LAB）是在粘膜水平递送抗原的有吸引力的工具。考虑到转基因LAB的安全性，我们用完全来自LAB的DNA构建了食品级分泌载体（pALRc或pALRb），包括复制子、启动子、信号肽和选择标记丙氨酸消旋酶基因（alr）。为了评估该系统的可行性，使用来自金黄色葡萄球菌的核酸酶基因（nuc）作为报告基因，在乳酸乳球菌和干酪乳杆菌中表达。随后，通过蛋白质印迹分析证实了ETEC的菌毛粘附素FaeG的细胞外分泌。这些结果表明，这种食品级表达系统有潜力作为安全使用转基因 LAB 来开发 ETEC 感染疫苗的输送工具。
Enterotoxigenic Escherichia coli (ETEC) infection is the major cause of diarrhea in neonatal piglets. The fimbriae as colonizing factor in the pathogenesis of ETEC constitute a primary target for vaccination against ETEC. Lactic acid bacteria (LAB) are attractive tools to deliver antigens at the mucosal level. With the safety of genetically modified LAB in mind, a food-grade secretion vector (pALRc or pALRb) was constructed with DNA entirely from LAB, including the replicon, promoter, signal peptide, and selection marker alanine racemase gene (alr). To evaluate the feasibility of the system, the nuclease gene (nuc) from Staphylococcus aureus was used as a reporter to be expressed in both Lactococcus lactis and Lactobacillus casei. Subsequently, the extracellular secretion of the fimbrial adhesin FaeG of ETEC was confirmed by Western blot analysis. These results showed that this food-grade expression system has potential as the delivery vehicle for the safe use of genetically modified LAB for the development of vaccines against ETEC infection.