title: DNA shuffling approach for recombinant polyvalent OmpAs against V. alginolyticus and E. tarda infections.
abstract: Molecular breeding via DNA shuffling directs the evolution of vaccines with desired traits. In the present study, polyvalent OmpA vaccines were generated by DNA shuffling of five ompA genes from four species of bacteria Vibrio parahaemolyticus, V. alginolyticus, Edwardsiella tarda and Escherichia coli. First, a new hybrid OmpA was constructed using VA0764 primers and used for construction of a prokaryotic expressing library PompAs-FV containing 84 ompAs, which were validated by PCR and SDS/PAGE. Then, the 84 ompAs were used to construct a eukaryotic expressing library EompAs-FV for preparing DNA vaccines. Third, extracellular bacterium V. alginolyticus challenge post active immunization using these DNA vaccines was carried out to identify genes with high immunoprotection. Among the 84 ompAs, 17 showed higher or equal immune protection against infection caused by V. alginolyticus than control VA0764. Finally, immune protection against infection caused by intracellular bacterium Edwardsiella tarda was assessed further using the top seven out of the 17 ompAs. This led to identification of three efficient polyvalent vaccines against infections caused by the extracellular bacterium V. alginolyticus and intracellular bacterium E. tarda. In addition, we sequenced genes for understanding mechanisms of the polyvalent vaccines, but association of immune protection with mutation of gene and amino acids is not determined. These results indicate that DNA shuffling is an efficient way to develop polyvalent vaccines against microbial infections.
authors.affiliation: Center for Proteomics and Metabolomics, State Key Laboratory of Bio-Control, MOE Key Lab Aquatic Food Safety, Guangdong Province Key Laboratory for Pharmaceutical Functional Genes, School of Life Sciences, Sun Yat-sen University, University City, Guangzhou 510006, People's Republic of China. Electronic address: lihui32@sysu.edu.cn.
authors.forename: Hui
authors.initials: H
authors.lastname: Li
authors.affiliation: Center for Proteomics and Metabolomics, State Key Laboratory of Bio-Control, MOE Key Lab Aquatic Food Safety, Guangdong Province Key Laboratory for Pharmaceutical Functional Genes, School of Life Sciences, Sun Yat-sen University, University City, Guangzhou 510006, People's Republic of China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Chu
authors.affiliation: Center for Proteomics and Metabolomics, State Key Laboratory of Bio-Control, MOE Key Lab Aquatic Food Safety, Guangdong Province Key Laboratory for Pharmaceutical Functional Genes, School of Life Sciences, Sun Yat-sen University, University City, Guangzhou 510006, People's Republic of China.
authors.forename: Bo
authors.initials: B
authors.lastname: Peng
authors.affiliation: Center for Proteomics and Metabolomics, State Key Laboratory of Bio-Control, MOE Key Lab Aquatic Food Safety, Guangdong Province Key Laboratory for Pharmaceutical Functional Genes, School of Life Sciences, Sun Yat-sen University, University City, Guangzhou 510006, People's Republic of China.
authors.forename: Xuan-Xian
authors.initials: XX
authors.lastname: Peng
mesh_terms: D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D001428:Bacterial Vaccines / Q000276:immunology*; D032621:DNA Shuffling / Q000662:veterinary; D020609:Edwardsiella tarda / Q000276:immunology*; D004756:Enterobacteriaceae Infections / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control / Q000662:veterinary; D004926:Escherichia coli / Q000276:immunology*; D004927:Escherichia coli Infections / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control / Q000662:veterinary; D022361:Escherichia coli Vaccines / Q000276:immunology; D005393:Fish Diseases / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control*; D014733:Vibrio / Q000276:immunology*; D014735:Vibrio Infections / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control / Q000662:veterinary; D044149:Vibrio alginolyticus / Q000276:immunology; D014736:Vibrio parahaemolyticus / Q000276:immunology; D015027:Zebrafish
chemical_list: D001425:Bacterial Outer Membrane Proteins; D001428:Bacterial Vaccines; D022361:Escherichia coli Vaccines; C078167:OMPA outer membrane proteins
keywords: DNA shuffling; E. tarda; Fish; Polyvalent vaccine; V. alginolyticus