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我们先前已经将人工合成的恶性疟原虫杂合１１３肽基因克隆到表达载体ｐＷＲ４５０－１中，获得了重组质粒ｐＷＲＡＢ。现将ｎＷＲＡＢ先转化到ＬＢ５０００修饰后，再转化到疫苗菌ＳＬ３２６１，得到重组减毒鼠伤寒沙门氏菌ＳＬ３２６１（ｐＷＲＡＢ）。ｐＷＲＡＢ在ＳＬ３２６１中以β－半乳糖苷酶一杂合多肽抗原ＡＢ融合蛋白（ＧＺ－Ａｍ形式表达，表达量约１３．３％。免疫双扩散发现从ＳＬ３２６１（ｐＷＲＡＢ）中纯化的ＧＺ－ＡＢ可与抗ＧＺ－ＡＢ抗体反应，滴度为１：１６。蛋白质印渍技术（Ｗｅｓｔｅｒｎｂｌｏｔ）显示在ＧＺ－ＡＢ相应位置处出现特异条带。上述结果初步说明ＳＬ３２６１表达的ＧＺ－ＡＢ具有抗原性。连续传代ＳＬ３２６１（ｐＷＲＡＢ）未见质粒ｐＷＲＡＢ丢失及明显的毒性，为恶性疟口服活疫苗的制备打下了基础。
We have previously cloned the synthetic Plasmodium falciparum heterozygous 113 peptide gene into the expression vector pWR450-1 to obtain the recombinant plasmid pWRAB. Now nWRAB first transformed into LB5000 modification, and then transformed into the vaccine strain SL3261, resulting in recombinant attenuated Salmonella typhimurium SL3261 (pWRAB). pWRAB was expressed in the form of GZ-Am with a β-galactosidase-heterozygous polypeptide antigen AB expression level of about 13.3% in SL3261. Immuno-double diffusion revealed that GZ-AB purified from SL3261 (pWRAB) Reacted with anti-GZ-AB antibody with a titer of 1:16 Western blot showed that a specific band appeared at the corresponding position of GZ-AB.The above results preliminarily showed that GZ-AB expressed by SL3261 has antigenicity Passage SL3261 (pWRAB) no plasmid pWRAB loss and significant toxicity, laid the foundation for the preparation of live oral vaccine against malignant malaria.