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出 处:《中国免疫学杂志》1995年第2期68-69,共2页Chinese Journal of Immunology
摘 要:通过Plkc噬菌体将Tn10插入灭活的aroD基因片段导入双价有毒痢疾菌FS-15中,构建了该株的芳香族氨基酸营养缺陷减毒株,并通过Bochner培养基去除Tn10所携带的四环素抗性,经原位杂交从中检测到aroD基因缺失株FS-5441。该株稳定表达福氏及宋内氏双价O抗原,Western-blot显示该株表达Ipa蛋白,具有穿入HeLa细胞的能力,穿入率为19%。营养缺陷表型的回复突变率小于10 ̄(-11)。小白鼠毒力试验证明安全无毒,免疫小白鼠后对福氏及宋内氏毒株的攻击具有保护活性。In this study,we constructed 10 attenuated strains by intruducing Tn10 inactived aroDgene into FS-15,a virulent bivalent hybrid strain of S. flexneri 2a and S.sonnei constructed inour laboratory in 1988,and then by using a modified bochner’s medium,a series of tetracycline-sensitive mutants were obtained.One mutant of them FS-5441 was demonstrated aroD genedeletion with a colony-hybridation with specific aroD gene probe,FS-5441 was found to invadeHela cells in vitro and negative in Sereny test.It expressed stably the somatic antigen of S.flexneri 2a and S.sonnei and exhibited significant protection for immunnized mice against thechallenge of S.flexeneri 2a and S.sonnei virulent strains.
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