Almeida Juliana F, Mariat Denis, Azevedo Vasco, Miyoshi Anderson, de Moreno de LeBlanc Alejandra, Del Carmen Silvina, Martin Rebeca, Langella Philippe, LeBlanc Jean-Guy, Chatel Jean-Marc
BMC Microbiol. 2014 Sep 25;14:248. doi: 10.1186/s12866-014-0248-9.
Fibronectin Binding Protein A (FnBPA) is an invasin from Staphylococcus aureus that allows this pathogen to internalize into eukaryote cells. It was previously demonstrated that recombinant Lactococcus lactis expressing FnBPA were invasive and able to transfer a plasmid to eukaryotic cells in vitro and in vivo. In this study, the invasivity of recombinant strains of Lactococcus lactis that express FnBPA under the control of its constitutive promoter or driven by the strong nisin inducible expression system (NICE) were studied.
It was demonstrated that the nisA promoter allows an increase of FnBPA expression on the surface of Lactococcus lactis surface, as shown by flow cytometry, which subsequently enhanced internalization and plasmid transfer properties in vitro in Caco2 cells and Bone Marrow Dendritic Cells. In vivo, the use of nisA promoter increase the plasmid transfer in cells of both the small and large intestine of mice.
FnBPA expression at the surface of recombinant L. lactis is positively correlated to internalization and DNA transfer properties. The recombinant strains of L. lactis that expresses FnBPA under the control of the nisin inducible expression system could thus be considered as an improved tool in the field of DNA transfer.
纤连蛋白结合蛋白A(FnBPA)是金黄色葡萄球菌的一种侵袭素,可使该病原体内化进入真核细胞。先前已证明,表达FnBPA的重组乳酸乳球菌具有侵袭性,并且能够在体外和体内将质粒转移至真核细胞。在本研究中,对在其组成型启动子控制下或由强乳链菌肽诱导表达系统(NICE)驱动表达FnBPA的重组乳酸乳球菌菌株的侵袭性进行了研究。
流式细胞术显示,nisA启动子可使FnBPA在乳酸乳球菌表面的表达增加,随后增强了其在体外对Caco2细胞和骨髓树突状细胞的内化及质粒转移特性。在体内,使用nisA启动子可增加小鼠小肠和大肠细胞中的质粒转移。
重组乳酸乳球菌表面的FnBPA表达与内化及DNA转移特性呈正相关。因此,在乳链菌肽诱导表达系统控制下表达FnBPA的重组乳酸乳球菌菌株可被视为DNA转移领域的一种改良工具。