论文部分内容阅读
目的检测含恶性疟原虫裂殖子表面蛋白１基因片断的重组减毒鼠伤寒杆菌Ｘ４０６４（ｐＱＥＭ１）的入侵宿主能力，所含质粒的稳定性和再表达能力。方法用含恶性疟原虫裂殖子表面蛋白１第１号基因片段的重组减毒鼠伤寒杆菌Ｘ４０６４（ｐＱＥＭ１）灌胃接种ＢＡＬＢ／ｃ小鼠，用平板培养法、质粒酶切和免疫印迹反应，鉴定从小鼠组织中分离到的该重组菌，测定其表达能力，及在小鼠组织中的消长。结果从小鼠组织中分离到的减毒鼠伤寒杆菌Ｘ４０６４株含有重组的ｐＱＥＭ１质粒；减毒鼠伤寒杆菌Ｘ４０６４（ｐＱＥＭ１）体外表达了恶性疟原虫裂殖子表面蛋白１第１号片断；Ｘ４０６４（ｐＱＥＭ１）在小鼠组织中的消长曲线与Ｘ４０６４的消长曲线基本一致。结论重组ｐＱＥＭ１质粒能稳定地存在于减毒鼠伤寒杆菌Ｘ４０６４株中，不妨碍其入侵宿主细胞，从小鼠组织中分离到的Ｘ４０６４（ｐＱＥＭ１）依然具有表达Ｍ１蛋白的能力。
Objective To detect the host capacity of invasion, the stability and re-expression of plasmids contained in the attenuated Salmonella typhimurium X4064 (pQEM1) containing the fragment of merozoite surface protein 1 gene of Plasmodium falciparum. Methods BALB / c mice were inoculated intragastrically with the recombinant attenuated Salmonella typhimurium X4064 (pQEM1) containing Plasmodium falciparum merozoite surface protein 1 gene fragment. The cells were stained with the method of plate culture, plasmid digestion and western blotting, The recombinant strain isolated from the mouse tissues was identified, its expression ability and its growth in mouse tissues were determined. Results The attenuated Salmonella typhimurium strain X4064 isolated from mouse tissues contained the recombinant plasmid pQEM1. The attenuated Salmonella typhimurium X4064 (pQEM1) expressed the first fragment of Plasmodium falciparum merozoite surface protein 1 in vitro. X4064 (pQEM1 ) In the mouse tissue growth curve and X4064’s growth curve is basically the same. Conclusion The recombinant plasmid pQEM1 can stably exist in attenuated Salmonella typhimurium X4064 strain without affecting its invasion into host cells. X4064 (pQEM1) isolated from mouse tissues still has the ability of expressing M1 protein.