title: Construction of a food-grade cell surface display system for Lactobacillus casei.
abstract: In this study, a food-grade cell surface display host/vector system for Lactobacillus casei was constructed. The food-grade host L. casei Q-5 was a lactose-deficient derivative of L. casei ATCC 334 obtained by plasmid elimination. The food-grade cell surface display vector was constructed based on safe DNA elements from lactic acid bacteria containing the following: pSH71 replicon from Lactococcus lactis, lactose metabolism genes from L. casei ATCC 334 as complementation markers, and surface layer protein gene from Lactobacillus acidophilus ATCC 4356 for cell surface display. The feasibility of the new host/vector system was verified by the expression of green fluorescent protein (GFP) on L. casei. Laser scanning confocal microscopy and immunofluorescence analysis using anti-GFP antibody confirmed that GFP was anchored on the surface of the recombinant cells. The stability of recombinant L. casei cells in artificial gastrointestinal conditions was verified, which is beneficial for oral vaccination applications. These results indicate that the food-grade host/vector system can be an excellent antigen delivery vehicle in oral vaccine construction.
authors.affiliation: College of Pharmacy, Binzhou Medical University, Yantai 264003, People's Republic of China. Electronic address: qinjysdu@gmail.com.
authors.forename: Jiayang
authors.initials: J
authors.lastname: Qin
authors.affiliation: College of Pharmacy, Binzhou Medical University, Yantai 264003, People's Republic of China. Electronic address: wangxiuwen0822@163.com.
authors.forename: Xiuwen
authors.initials: X
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, People's Republic of China. Electronic address: kongjian@sdu.edu.cn.
authors.forename: Jian
authors.initials: J
authors.lastname: Kong
authors.affiliation: State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, People's Republic of China. Electronic address: macq@sdu.edu.cn.
authors.forename: Cuiqing
authors.initials: C
authors.lastname: Ma
authors.affiliation: State Key Laboratory of Microbial Metabolism, and School of Life Sciences & Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China. Electronic address: pingxu@sjtu.edu.cn.
authors.forename: Ping
authors.initials: P
authors.lastname: Xu
mesh_terms: D061505:Cell Surface Display Techniques; D005455:Fluorescent Antibody Technique; D005516:Food Microbiology; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D007780:Lacticaseibacillus casei / Q000235:genetics* / Q000378:metabolism*; D008565:Membrane Proteins / Q000032:analysis / Q000235:genetics; D018613:Microscopy, Confocal; D010957:Plasmids; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D008565:Membrane Proteins; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins
keywords: Food-grade; Lactobacillus casei; Plasmid elimination; Surface display