BACILLUS-SUBTILIS EXPRESSING A HEMOLYSIN GENE FROM LISTERIA-MONOCYTOGENES CAN GROW IN MAMMALIAN-CELLS
BACILLUS-SUBTILIS EXPRESSING A HEMOLYSIN GENE FROM LISTERIA-MONOCYTOGENES CAN GROW IN MAMMALIAN-CELLS
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DOI：
10.1038/345175a0
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发表时间：
1990-05-10
期刊：
影响因子：
64.8
通讯作者：
PORTNOY, DA
中科院分区：
文献类型：
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作者：
BIELECKI, J;YOUNGMAN, P;PORTNOY, DA
细胞内寄生虫可分为寄居在宿主液泡内的寄生虫和直接在宿主细胞质中生长的寄生虫1。后一组的成员，包括立克次体 2、志贺氏菌 3、克氏锥虫 4 和单核细胞增生李斯特氏菌 5,6，具有与进入宿主细胞质的能力相关的溶血活性 5-7。因此L的突变体。缺乏成孔溶血素李斯特氏菌溶血素 O 的单核细胞增生症不会从内体区室逃逸 5,6，因此无法在细胞质中建立5,8,9。为了研究李斯特氏菌溶血素 O 的作用，我们克隆了李斯特菌溶血素 O 的结构基因。单核细胞增生溶血素 hlyA 在 IPTG 诱导型启动子的控制下转化为枯草芽孢杆菌的无孢子突变体10。被巨噬细胞样细胞系 J774 内化后，溶血 B。枯草芽孢杆菌破坏吞噬体膜并在巨噬细胞细胞质内快速生长。这些结果表明，单个基因产物足以将常见的土壤细菌转化为可以在哺乳动物细胞的细胞质中生长的寄生虫。
INTRACELLULAR parasites can be classified into those that reside within a host vacuole and those which grow directly in the host cytoplasm1. Members of the latter group, which includesRickettsia2,Shigellae3,Trypanosoma cruzi4, andListeria monocytogenes5,6, possess haemolytic activity associated with the ability to enter the host cytoplasm5–7. Therefore mutants ofL. monocytogeneslacking a pore-forming haemolysin, listeriolysin O, do not escape from the endosomal compartment5,6and consequently fail to become established in the cytoplasm5,8,9. To examine the role of listeriolysin O, we cloned the structural gene for theL. monocytogeneshaemolysin,hlyA, into an asporogenic mutant ofBacillus subtilisunder the control of an IPTG-inducible promoter10. After being internalized by the macrophage-like cell line J774, haemolyticB. subtilisdisrupted the phagosomal membrane and grew rapidly within the macrophage cytoplasm. These results show that a single gene product is sufficient to convert a common soil bacterium into a parasite that can grow in the cytoplasm of a mammalian cell.