日本血吸虫SjRAD23基因的克隆表达及基因重组抗原的免疫保护效果
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

国家自然科学基金 (Nos. 31172315, 81271871),上海市科技发展基金 (No. 12140902700),中国农业科学院院长基金 (No. 2013JB18),中国博士后科学基金 (No. 2012M510630) 资助。


Cloning, expression and protective efficacy evaluation of radiation sensitive protein 23 (RAD23) from Schistosoma japonicum
Author:
Affiliation:

Fund Project:

National Natural Science Foundation of China (Nos. 31172315, 81271871), Science and Technology Commission of Shanghai Municipality (No. 12140902700), Basic Scientific Research Operation Cost of State-level Public Welfare Scientific Research Courtyard (No. 2013JB18), China Postdoctoral Science Foundation (No. 2012M510630)

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    辐射敏感蛋白23具有核苷酸切除修复功能,在泛素蛋白酶体途径中起到重要作用。本研究利用PCR技术克隆了日本血吸虫辐射敏感蛋白23 (SjRAD23) 编码的cDNA序列,成功获得SjRAD23的基因序列,其ORF为1 053 bp。构建SjRAD23基因重组表达质粒pET28a(+)-SjRAD23,并在大肠杆菌BL21中成功诱导表达,重组蛋白在上清和沉淀中都有存在。利用免疫组化技术检测该蛋白在虫体的分布情况,该蛋白广泛分布在日本血吸虫虫体被膜。用重组蛋白免疫BALB/c小鼠后,免疫小鼠血清中检测到较高水平的特异性IgG、IgG1和IgG2a。Western blotting分析显示重组蛋白能够被日本血吸虫成虫可溶性抗原免疫小鼠血清所识别。用重组蛋白rSjRAD23免疫结果与206佐剂对照组比较,rSjRAD23在BALB/c小鼠中诱导了35.94%减虫率,40.59%肝脏减卵率。结果表明SjRAD23具有作为疫苗候选分子的潜力。

    Abstract:

    Radiation sensitive protein 23 (RAD23) is a nucleotide excision repair (NER) protein that plays an important role in Ubiquitin-proteasome pathway (UPP). Schistosoma japonicum radiation sensitive protein23 (SjRAD23) cDNA sequences were amplified by PCR and cloned into pET28a (+) vector to construct recombinant expression plasmid pET28a (+)-SjRAD23. The recombinant protein was expressed as both inclusion bodies and the supernatant in Escherichia coli BL21 (DE3) cell. Immunofluorescence observation shows that SjRAD23 was mainly distributed on the tegument surface of the worms. ELISA assay reveals that specific IgG, IgG1 and IgG2a antibodies could be detected in the sera of rSjRAD23 immunized mice. Western blotting analysis shows that the recombinant SjRAD23 could be recognized by serum specific to soluble adult worm antigen of S. japonicum. BALB/c mice vaccinated with rSjRAD23 combined with 206 adjuvant revealed 35.94% worm reduction and 40.59% liver egg reduction when compared with that of the adjuvant control group .This study suggests that rSjRAD23 has the potential as a vaccine against schistosome infection.

    参考文献
    相似文献
    引证文献
引用本文

李长健,张旻,洪炀,韩艳辉,曹晓丹,韩宏晓,傅志强,朱传刚,陆珂,李浩,林矫矫. 日本血吸虫SjRAD23基因的克隆表达及基因重组抗原的免疫保护效果[J]. 生物工程学报, 2014, 30(11): 1669-1678

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2014-02-19
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期: 2014-10-31
  • 出版日期:
您是第位访问者
生物工程学报 ® 2024 版权所有

通信地址:中国科学院微生物研究所    邮编:100101

电话:010-64807509   E-mail:cjb@im.ac.cn

技术支持:北京勤云科技发展有限公司