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日本血吸虫丝氨酸苏氨酸蛋白磷酸酶基因的表达
及其免疫保护试验
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姚利晓1,2，孙安国1,2，傅志强1，刘金明1，苑纯秀1， 蔡幼民1，林矫矫1
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(1. 中国农业科学院 上海家畜寄生虫病研究所，上海200232；
2. 中国农业科学院 研究生院，北京100081 )
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摘要：
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为研究日本血吸虫丝氨酸苏氨酸蛋白磷酸酶(SjPP)的免疫功能，构建了原核表达重组质粒pET28a(+)SjPP，转化大肠埃希氏菌BL21(DE3)进行诱导表达，并用重组蛋白进行了小鼠免疫保护试验，免疫剂量为每次20μg/只，共免疫3次。结果表明，pET28a(+)SjPP/BL21(DE3)在01mmol/L IPTG诱导2h时，每1g菌体可产生178mg以包涵体形式存在的重组蛋白；重组蛋白免疫小鼠后诱导产生了高效价的血清特异性IgG抗体，并获得了部分减虫率、肝组织减卵率和显著的粪便减卵率，具有一定的免疫保护作用。 |
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关键词：
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日本血吸虫；丝氨酸苏氨酸蛋白磷酸酶；蛋白表达；免疫保护作用 |
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中图分类号：S 852.735: Q 786 文献标识码：
A 文章编号：1673-4696(2006)02-0122-05 |
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Expression and protective immunity of serinethreonine specific
protein phosphatase SjPP of Schistosoma japonicum |
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YAO Lixiao1,2，SUN
Anguo1,2, FU Zhiqiang1, LIU
Jinming1,
YUAN Chunxiu1，CAI Youmin1, LIN
Jiaojiao1 |
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（1.Shanghai Institute of Animal Parasitology, Chinese Academy of Agricultural Sciences, Shanghai200232, China;
2. School of Graduate, Chinese Academy of Agricultural Sciences, Beijing100081, China) |
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Abstract: A serinethreonine specific protein phosphatase gene of Schistosoma japonicum, SjPP, was cloned into pET28a(+) to construct a recombinant plasmid pET28a(+)SjPP. The plasmid pET28a(+)SjPP in E. coli strain BL21(DE3) was induced with 01mmol/L of IPTG and expressed in inclusion bodies. Production of the recombinant protein rSjPP was 178mg per gram of bacteria after induction for 2h. The purified recombinant proteins were used to vaccinate BALB/c mice to evaluate immunoprotective efficacy. The mice in vaccination group were immunized with 20μg rSjPP emulsified with Freund′s complete adjuvant(FCA), and those in adjuvant or blank control groups were injected with FCA/FIA or PBS, respectively on week 0, 2, 4. The mice were challenged with cercariae on week 2 after the final injection, and were sacrificed on day 42 after challenge. High titre specific antirSjPP antibody IgG was observed in the sera of vaccinated mice, and partial worm reduction, hepatic tissue egg reduction, and significant faecal egg reduction were obtained in vaccinated group, when comparied with that in the blank control group. |
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Key words: Schistosoma japonicum; serinethreonine specific protein phosphatase; protein expression; immunoprotection |
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