[1]赵玉霞)△,杨国俊),章涵),等.幽门螺杆菌UreBOmp11融合蛋白的表达、纯化与免疫学活性检测[J].郑州大学学报(医学版),2009,(01):125-128.
　ZHAO Yuxia),YANG Guojun),ZHANG Han),et al.Expression and purification of UreBOmp11 fusion protein of Helicobacter pylori and its immunocompetence[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2009,(01):125-128.
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幽门螺杆菌UreBOmp11融合蛋白的表达、纯化与免疫学活性检测()
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2009年01期
页码:
125-128
栏目:
论著
出版日期:
2009-01-01

文章信息/Info

Title:
Expression and purification of UreBOmp11 fusion protein of Helicobacter pylori and its immunocompetence
作者:
赵玉霞1)△; 杨国俊2); 章涵1); 段广才3; 4) #; 郗园林4)
1)信阳职业技术学院医学系信阳4640002）河南职工医学院郑州4500523）河南省分子医学重点学科开放实验室郑州4500524)郑州大学公共卫生学院流行病学教研室郑州450001
Author(s):
ZHAO Yuxia1); YANG Guojun2); ZHANG Han1); DUAN Guangcai3; 4); XI Yuanlin4)
1)Department of Clinical Medicine,Xinyang Vocational and Technical College,Xinyang 4640002) Henan Medical College for Staff and Work,Zhengzhou 4500523)Department of Epidemiology,College of Public Health,Zhengzhou University,Zhengzhou 4500524)Key Open Laboratory of Molecular Medcine of Henan Province,Zhengzhou 450001
关键词:
幽门螺杆菌; UreBOmp11; 免疫学
Keywords:
Helicobacter pylori;  UreBOmp11;  protein expression;  immunocompetence
分类号:
R183
摘要:
在大肠杆菌TB1(pMALc2X)中表达幽门螺杆菌（Hp）UreBOmp11融合蛋白，并探索其免疫学活性，为Hp基因工程疫苗的研制奠定基础。方法：以重组质粒pET30a(+)ureBomp11为模板获得融合基因ureBomp11片段，并插入到原核表达载体pMALc2X中进行融合蛋白的表达，用Amylose亲和层析法纯化融合蛋白，纯化的融合蛋白辅以免疫佐剂皮下免疫小鼠，蛋白印记法对免疫小鼠血清进行检测。结果：特异PCR法和酶切鉴定证实融合基因ureBomp11克隆入表达载体pMALc2X中；重组菌TB1(pMALureBomp11)经诱导获得了高效表达的MBPUreBOmp11融合蛋白，该融合蛋白可以被Hp免疫小鼠血清的相应抗体所识别，纯化后的融合蛋白纯度达90%以上。通过大肠杆菌抗原吸收法纯化免疫小鼠血清后，与纯化的融合蛋白进行杂交，结果显示在134000处出现特异杂交带，融合蛋白具有良好的免疫学活性。结论：成功构建了HpMELHP27融合蛋白UreBOmp11的重组疫苗候选株TB1(pMALureBomp11)，为Hp蛋白质疫苗和核酸疫苗的研制奠定了良好的基础。
Abstract:
To express and purify the fusion protein UreBOmp11 and to determine its immunocompetence.Methods:the fusion gene UreBOmp11 were amplified from recombinant plasmid pET30a(+)UreBOmp11 and cloned into the fusion expression vector pMALc2X. then the recombinant UreBOmp11 fusion protein was expressed and indentified by SDSPAGE and Western Blot analysis. Then the fusion protein was purified by MBP affinity chromatography and the purified fusion protein was immunized to mice. the immunized mice sera were analyzed by Western Blot with purified fusion protein.Results: The UreBOmp11 fusion gene was correctly insected into pMALc2X and confirmed by Enzyme digestion and PCR; Results in SDSPAGE and optical density scanning demonstrated that this fusion protein MBPUreBOmp11 was expressed in the recombinant strain of E.coli TB1(pMALUreBOmp11). The fusion protein SDSPAGE and optical density scanning demonstrated that this fusion protein MBPUreBOmp11 was expressed in the recombinant strain of E.coli TB1(pMALUreBOmp11). The fusion protein UreBOmp11 was recognized by the mice sera immunized by H.pylori，the purity of fusion protein was 90% after purification. The fusion protein purified could be recognized by corresponding antibody of mice sera immunized by this fusion piotein, this fusion protein has strong immunocompetence.Conclusion: The prokaryotic expression system TB1 (pMALc2XUreBOmp11) was successfully constructed. The results obtained lay the foundation for research on development of protein and DNA vaccine of H.pylori.

参考文献/References:

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备注/Memo

备注/Memo:
△女，1980年生，硕士，助教，研究方向：幽门螺杆菌基因工程疫苗的研究，Email:zhaoyuxia888@126.com #通讯作者，男，1958年生，博士，教授，研究方向：分子流行病学，Email:gcduan@public.zz.ha.cn
更新日期/Last Update: 2010-05-18