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日本血吸虫SjPRMT1基因的克隆、表达及表达产物的免疫保护效果分析
  • ISSN号:0578-1752
  • 期刊名称:《中国农业科学》
  • 时间:0
  • 分类:S858.28[农业科学—临床兽医学;农业科学—兽医学;农业科学—畜牧兽医] Q959.4[生物学—动物学]
  • 作者机构:[1]中国农业科学院上海兽医研究所/农业部动物寄生虫学重点开放实验室,上海200241, [2]南京农业大学动物医学院,南京210095
  • 相关基金:国家自然科学基金(30671581)、“973”课题(2007CB513108)、“863”计划(2006AA10A207)
中文摘要:

【目的】克隆和表达臼本血吸虫精氨酸甲基转移酶1(SjPRMT1)编码基因cDNA,分析其在日本血吸虫不同发育阶段虫体的表达情况,评估该重组抗原在小鼠体内诱导的抗血吸虫免疫保护效果。【方法】从实验室构建的7d童虫消减cDNA文库中,获得一个EST序列,经序列测定和生物信息学分析命名为SJPRMT1,以彤cDNA为模板,获得其全长cDNA。荧光实时定量PCR分析该基因在童虫和成虫的表达情况。以pET28a(+)为载体构建重组表达质粒,Westernblotting检测重组蛋白的抗原性-9免疫原性,利用重组抗原免疫小鼠评估其免疫保护效果。【结果】获得了SjpRMT1编码基因的全长eDNA,开放阅读框为660bp,编码219个氨基酸。荧光实时定量PCR分析该基因在18d童虫高表达,13和23d次之。获得了SjPRMT1重组蛋白,其具有良好的抗原性和免疫原性。在小鼠免疫试验中,与空白对照组比较,免疫组小鼠获得35.07%的减虫率和48.66%的肝脏减卵率。【结论】获得了日本血吸虫童虫期高表达的SJPRMT1基因的全长eDNA,成功构建了pET28a(+)-SjPRMT1重组质粒,该重组抗原在小鼠体内诱导产生了部分免疫保护效果。

英文摘要:

[ Objective ] The present study was intend to clone a full length cDNA encoding protein arginine methyltransferase 1 in Schistosoma japonicum, and further access its immunoprotection in mice for schistosomiasis. [ Method ] A full-length cDNA clone encoding a protein arginine methyltransferase was obtained from schistosomula cDNA enriched library, named S.jPRMT1 based on bioinformatics analysis. The expression profiles of SjPRMT1 was determined by real-time PCR using the template cDNAs isolated from 7, 13, 18, 23, 32 and 42 days parasites. The open reading frame was then subcloned into a pET28a(+) vector and transformed into competent E.coil/BL21. The recombinant protein was purified and then the immune characters of antigen were accessed by Western blotting. [Result] Bioinformatics analysis indicated that the ORF of S.jPRMT1 is 660 base pairs, which is encoding 219 amino acids. Real-time PCR analysis revealed that there is a highest expression in 18 days schistosomula, and slightly lower in 13 days and 23 days parasites, suggesting that S.jPRMT1 has highly expression in schistosomula stage. The recombinant protein showed a good ability to induce antibodies in mice as examined by ELISA. Animal experiment indicated that the administration of recombinant PRMT1 protein in mice resulted in worm burden reduced by 35.07% and egg burden reduced by 48.66%. [Conclusion] A full-length cDNA differently expression in schistosomula was obtained. Results demonstrated that the recombinant PRMT1 protein could induce partial protection against schistosomiasis in mice.

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期刊信息
  • 《中国农业科学》
  • 中国科技核心期刊
  • 主管单位:中华人民共和国农业部
  • 主办单位:中国农业科学院 中国农学会
  • 主编:万建民
  • 地址:北京中关村南大街12号中国农业科学院图书馆楼4101-4103室
  • 邮编:100081
  • 邮箱:zgnykx@caas.cn
  • 电话:010-82109808 82106279
  • 国际标准刊号:ISSN:0578-1752
  • 国内统一刊号:ISSN:11-1328/S
  • 邮发代号:2-138
  • 获奖情况:
  • 中国期刊方阵“双高”期刊,第三届中国出版政府奖提名奖
  • 国内外数据库收录:
  • 美国化学文摘(网络版),英国农业与生物科学研究中心文摘,波兰哥白尼索引,英国动物学记录,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),英国食品科技文摘,中国北大核心期刊(2000版)
  • 被引量:85620