title: Expression of porcine respiratory and reproductive syndrome virus membrane-associated proteins in Listeria ivanovii via a genome site-specific integration and expression system.
abstract: Listeria ivanovii (LI) possesses a facultative intracellular life cycle in macrophages and non-professional phagocytes thus making it a potential novel vaccine vector. We have developed a genome site-specific integration system as a universal tool for constructing the live LI-based vaccines. Using this system, the porcine reproductive and respiratory syndrome virus (PRRSV) orf5 and orf6 genes, coding for PRRSV membrane-associated protein GP5 and M, respectively, were integrated into the LI genome. Genome PCR and sequencing results showed that the orf5 or orf6 gene had been integrated into the LI genome downstream of orfXYZ or mpl gene. Western blot analysis of the non-secreted and secreted bacterial protein samples showed the expression and secretion of GP5 or M protein from the recombinant LI strains. The secretion of the heterologous proteins is a key step for bacterial vaccine vectors to deliver the antigens to the immune system to stimulate antigen-specific immune responses. Therefore, this integration system may be a potential tool for constructing novel vaccines using live LI as vector.
authors.affiliation: Department of Medical Technology, West China School of Public Health, Sichuan University, Chengdu, PR China.
authors.forename: Chuan
authors.initials: C
authors.lastname: Wang
authors.forename: Fuqin
authors.initials: F
authors.lastname: Zhang
authors.forename: Jianglong
authors.initials: J
authors.lastname: Yang
authors.forename: Asma
authors.initials: A
authors.lastname: Khanniche
authors.forename: Hao
authors.initials: H
authors.lastname: Shen
mesh_terms: D000956:Antigens, Viral / Q000096:biosynthesis* / Q000235:genetics*; D015153:Blotting, Western; D004337:Drug Carriers; D015870:Gene Expression; D005822:Genetic Vectors; D008087:Listeria / Q000235:genetics* / Q000378:metabolism; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics*; D016133:Polymerase Chain Reaction; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D011995:Recombination, Genetic
chemical_list: D000956:Antigens, Viral; D004337:Drug Carriers; D008565:Membrane Proteins; D011994:Recombinant Proteins