Husseiny Mohamed I, Hensel Michael
Department of Microbiology and Immunology, Faculty of Pharmacy, Zagazig University, Zagazig, Egypt.
Microbiol Res. 2008;163(6):605-15. doi: 10.1016/j.micres.2006.10.003.
Attenuated live Salmonella enterica serovar Typhimurium is a versatile organism for the generation of live recombinant vaccines for mucosal immunization and various approaches were devised for the stable and efficient expressions of heterologous antigens by attenuated S. enterica strains. Phage lamda Red recombinase has recently been devised for gene replacements in S. enterica after introduction of PCR products as a one-step deletion approach and FLP-mediated recombination allows the subsequent removal of antibiotic resistance markers. As an extension of this method, we have developed an approach that allows the sequential integration of multiple recombinant expression cassettes for heterologous antigens into the chromosome of S. enterica. We observed the stable expression of model antigens without selective pressure. In addition, the method allows the simultaneous generation of double-attenuating mutations by gene deletions. This approach allows the rapid and efficient construction of recombinant Salmonella strains as vaccine carriers.
减毒活肠炎沙门氏菌鼠伤寒血清型是一种用于生产黏膜免疫活重组疫苗的多功能生物体，并且已经设计出各种方法来通过减毒肠炎沙门氏菌菌株稳定高效地表达异源抗原。噬菌体λ Red重组酶最近被设计用于在引入PCR产物后在肠炎沙门氏菌中进行基因替换，这是一种一步缺失方法，而FLP介导的重组允许随后去除抗生素抗性标记。作为该方法的扩展，我们开发了一种方法，该方法允许将多个用于异源抗原的重组表达盒顺序整合到肠炎沙门氏菌的染色体中。我们观察到在没有选择压力的情况下模型抗原的稳定表达。此外，该方法允许通过基因缺失同时产生双重减毒突变。这种方法允许快速高效地构建作为疫苗载体的重组沙门氏菌菌株。
