日本血吸虫新基因sjf4的克隆、表达及功能分析

Liang Zhang, Guo-Feng Cheng, Zhi-Qiang Fu, Ya-Mei Jin, Xin-Gang Feng, Chun-Xiu Yuan, You-Min Cai, Jiao-Jiao Lin
{"title":"日本血吸虫新基因sjf4的克隆、表达及功能分析","authors":"Liang Zhang,&nbsp;Guo-Feng Cheng,&nbsp;Zhi-Qiang Fu,&nbsp;Ya-Mei Jin,&nbsp;Xin-Gang Feng,&nbsp;Chun-Xiu Yuan,&nbsp;You-Min Cai,&nbsp;Jiao-Jiao Lin","doi":"","DOIUrl":null,"url":null,"abstract":"<p><p>Based on the phenomenon of the natural anti-schistosomiasis in Microtus fortis, the sera from normal Microtus fortis were employed to immunoscreen the cDNA library of adult Schistosoma japonicum (Chinese strain), two positive clones were obtained, RACE technique was further applied to amplify one of the clones, and a cDNA fragment with an ORF was identified. Sequencing revealed that it was a novel gene of Schistosoma japonicum, and it was named SjMF4 (Schistosoma japonicum Microtus fortis 4). Then the structure and functional motifs of SjMF4 were analysed. The gene was subcloned into pET-28a(+) vector; the recombinant protein showed good antigenicity in Western blotting. The gene was further subcloned into eukaryotic expression vector pcDNA3 to construct the DNA vaccine containing SjMF4. Immune experiments in mice showed significant protection that the recombinant plasmid did induce 28.64%+/-3.82% worm reduction and 21.73%+/-3.98% egg reduction than controls against the Schistosoma japonicum cercaria challenge.</p>","PeriodicalId":21763,"journal":{"name":"Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2003-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"[Cloning, expressing and functional analysis of SjMF4, a novel Schistosoma japonicum gene].\",\"authors\":\"Liang Zhang,&nbsp;Guo-Feng Cheng,&nbsp;Zhi-Qiang Fu,&nbsp;Ya-Mei Jin,&nbsp;Xin-Gang Feng,&nbsp;Chun-Xiu Yuan,&nbsp;You-Min Cai,&nbsp;Jiao-Jiao Lin\",\"doi\":\"\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Based on the phenomenon of the natural anti-schistosomiasis in Microtus fortis, the sera from normal Microtus fortis were employed to immunoscreen the cDNA library of adult Schistosoma japonicum (Chinese strain), two positive clones were obtained, RACE technique was further applied to amplify one of the clones, and a cDNA fragment with an ORF was identified. Sequencing revealed that it was a novel gene of Schistosoma japonicum, and it was named SjMF4 (Schistosoma japonicum Microtus fortis 4). Then the structure and functional motifs of SjMF4 were analysed. The gene was subcloned into pET-28a(+) vector; the recombinant protein showed good antigenicity in Western blotting. The gene was further subcloned into eukaryotic expression vector pcDNA3 to construct the DNA vaccine containing SjMF4. Immune experiments in mice showed significant protection that the recombinant plasmid did induce 28.64%+/-3.82% worm reduction and 21.73%+/-3.98% egg reduction than controls against the Schistosoma japonicum cercaria challenge.</p>\",\"PeriodicalId\":21763,\"journal\":{\"name\":\"Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2003-12-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0

摘要

基于东方田鼠天然抗血吸虫病的现象,利用正常东方田鼠血清免疫筛选日本血吸虫(中国株)成虫cDNA文库,获得2个阳性克隆,应用RACE技术对其中1个克隆进行扩增,鉴定出具有ORF的cDNA片段。测序结果显示该基因为日本血吸虫新基因,并将其命名为sjf4 (Schistosoma japonicum Microtus fortis 4),分析了sjf4的结构和功能基序。将该基因亚克隆到pET-28a(+)载体中;Western blot结果显示重组蛋白具有良好的抗原性。将该基因亚克隆到真核表达载体pcDNA3中,构建含sjf4的DNA疫苗。小鼠免疫实验表明,与对照相比,重组质粒对日本血吸虫尾蚴攻击的虫减少率为28.64%+/-3.82%,卵减少率为21.73%+/-3.98%。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
[Cloning, expressing and functional analysis of SjMF4, a novel Schistosoma japonicum gene].

Based on the phenomenon of the natural anti-schistosomiasis in Microtus fortis, the sera from normal Microtus fortis were employed to immunoscreen the cDNA library of adult Schistosoma japonicum (Chinese strain), two positive clones were obtained, RACE technique was further applied to amplify one of the clones, and a cDNA fragment with an ORF was identified. Sequencing revealed that it was a novel gene of Schistosoma japonicum, and it was named SjMF4 (Schistosoma japonicum Microtus fortis 4). Then the structure and functional motifs of SjMF4 were analysed. The gene was subcloned into pET-28a(+) vector; the recombinant protein showed good antigenicity in Western blotting. The gene was further subcloned into eukaryotic expression vector pcDNA3 to construct the DNA vaccine containing SjMF4. Immune experiments in mice showed significant protection that the recombinant plasmid did induce 28.64%+/-3.82% worm reduction and 21.73%+/-3.98% egg reduction than controls against the Schistosoma japonicum cercaria challenge.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
自引率
0.00%
发文量
0
期刊最新文献
Analysis of gene expression in hepatitis B virus transfected cell line induced by interferon. [Cloning and expression of tumor necrosis factor (TNFalpha) cDNA from red seabream pagrus major]. [Effects of sodium selenite on telomerase activity and telomere length]. [Cloning, eukaryotic expression and function assay of recombinant leukemia inhibitory factor gene LIF]. [The immunopotentiation of human B lymphocyte stimulator C-terminal peptide].
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1