[1]葛迪,郭刚,毛旭虎,等.幽门螺杆菌UreB414-OMP18-LTB融合蛋白的构建及免疫原性初步研究[J].第三军医大学学报,2008,30(12):1107-1110.
 GE Di,GUO Gang,MAO Xu-hu,et al.Construction and immunogenicity of a fusion vaccine candidate of UreB-OMP18-LTB from H. pylori[J].J Third Mil Med Univ,2008,30(12):1107-1110.
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幽门螺杆菌UreB414-OMP18-LTB融合蛋白的构建及免疫原性初步研究(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
30卷
期数:
2008年第12期
页码:
1107-1110
栏目:
论著
出版日期:
2008-06-30

文章信息/Info

Title:
Construction and immunogenicity of a fusion vaccine candidate of UreB-OMP18-LTB from H. pylori
作者:
葛迪 郭刚毛旭虎张卫军邹全明
第三军医大学医学检验系临床微生物学及免疫学教研室,重庆市生物制药工程技术研究中心
Author(s):
GE Di GUO Gang MAO Xu-hu ZHANG Wei-jun ZOU Quan-ming
Laboratory of Clinic Microbiology and Immunology, College of Medical Laboratory, Third Military Medical University, Chongqing 400038, China
关键词:
幽门螺杆菌尿素酶外膜蛋白大肠埃希菌不耐热毒素B亚单位疫苗
Keywords:
Helicobacter pylori urease outer memberane protein18 E. coli heatlabile enterotoxin B subunit vaccine
分类号:
R378.99;R392.1;R394-33
文献标志码:
A
摘要:
目的  构建幽门螺杆菌(Helicobacter pylori,Hp) 尿素酶B亚单位活性片段(UreB414)、外膜蛋白(OMP18)及大肠埃希菌LTB融合疫苗并研究其抗原性。  方法  PCR扩增ureb414、omp18、ltb基因,分别克隆入pMD18-T载体,测序并鉴定方向后依次连接,构建融合基因UOL,然后将其亚克隆入原核表达载体pET-28a,转化入大肠埃希菌BL21,SDS-PAGE电泳进行表达,Western印迹检测表达蛋白的抗原性。  结果  PCR扩增分别获得约414、537 bp和320 bp产物,与GenBank中相关序列具有高度同源性,三者融合后获得一约1 200 bp目的基因。重组融合蛋白表达量可达细菌总蛋白的15%,免疫印迹检测结果显示该重组蛋白可为UreB、OMP18、LTB兔抗血清特异识别,具有良好的抗原性。  结论  成功构建了具有良好免疫原性的UreB-OMP18-LTB融合蛋白。
Abstract:
Objective    To construct a fusion vaccine of Helicobacter pylori (Hp) urease subunit B (UreB), outer membrane protein18 (OMP18) and E.coli heat-labile enterotoxin B subunit, summarized as UOL and study its antigenicity so as to lay a foundation to produce an H.pylori vaccine.     Methods    The UreB414, OMP18 and LTB genes were amplified by PCR, and cloned into pMD 18-T vector, and the products were identified and fused to abtain UOL. UOL was then subcloned into a procaryon expression vector pET-28a and transformed into E. coli cells BL21. Immunogenicity of the recombinant fusion protein was analyzed by Western blotting.     Results    The PCR fragments of ureb414, omp18 and ltb genes were about 414, 537 and 320 bp, which had high homology with those related sequences in GenBank. Recombinant protein amounted to 15% of the total bacterial protein. Western blot showed that the protein had a specific reaction with UreB, OMP18 and LTB antiserum.     Conclusion    Expression of the fusion protein of UOL in E. coli cells provides a basis to produce engineered vaccine against Hp.

参考文献/References:

葛迪, 郭刚,毛旭虎,等. 幽门螺杆菌UreB414-OMP18-LTB融合蛋白的构建及免疫原性初步研究第三军医大学学报,2008,30(12):1107-1110.

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更新日期/Last Update: 2008-06-13