Generation of safety enhanced Edwardsiella tarda ghost vaccine

Diseases of Aquatic Organisms

Dong Jin Lee¹  Sung Koo Kim¹  Eun Hye Lee¹  Kosuke Zenke¹  Ki Hong Kim¹  Se Ryun Kwon¹  Yoon Kwon Nam¹ 

关键词: Edwardsiella tarda;    Ghost bacteria;    Staphylococcal nuclease A;    Dual vector;    Safety enhancement;   

DOI  :  10.3354/dao01964

学科分类：生物科学（综合）

来源: Inter-Research

【 摘 要 】

ABSTRACT: A dual vector expressing the ghost-inducing PhiX174 lysis E gene and the bacterial DNA degrading staphylococcal nuclease A (SNA) gene was constructed to solve the problem of remnant antibiotic resistance genes and genomic DNA with intact pathogenic islands in the final product of Edwardsiella tarda ghosts (ETG). The SNA (devoid of secretion signal sequence and the nuclease B amino terminus sequence), fused with the 26 amino acid N-terminal sequence of the λ phage Cro gene, showed successful degradation of bacterial nucleic acids. Furthermore, the nuclease activity of SNA in E. tarda was enhanced by codon optimization of the SNA gene using site-directed mutagenesis. ETG were generated via coexpression of the SNA gene and lysis gene E under the control of each λP_R promoter. The ghost bacteria generation system we describe is advantageous as it allows the use of a single plasmid, improves safety and vaccine purity by limiting residual genetic content from the ghost bacteria, and reduces production costs through cheap means of induction that use only temperature shifts.

【 授权许可】

Unknown