• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>World Journal of Gastroenterology >Expression of Helicobacter pylori AlpA protein and its immunogenicity.
【24h】

Expression of Helicobacter pylori AlpA protein and its immunogenicity.

机译：幽门螺杆菌AlpA蛋白的表达及其免疫原性。

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

AIM: To construct a recombinant strain which expresses adhesin AlpA of Helicobacter pylori (H pylori) and to study the immunogenicity of adhesin AlpA. METHODS: Gene Ab, which was amplified from H pylori chromosomal DNA by PCR technique, was sequenced and the biological information was analyzed, and inserted into the Nco I and Not I restriction fragments of the expression vector pET-22b(+) using T4 DNA ligase. The resulting plasmid pET-AlpA was transformed into competent E.coli BL21(DE3) cells using ampicillin resistance for selection. Recombinant strains were incubated in 5 mL LB with 100 mug/mL ampicillin overnight at 37 degrees. Sonication of BL21(DE3)pET-22b(+)/AlpA was analyzed by Western blot to detect AlpA immunogenicity. RESULTS: The gene encoding AlpA protein was amplified by PCR with chromosomal DNA of H pylori Sydney strain (SS1) as templates. It revealed that AlpA DNA fragment amplified by PCR had approximately 1 500 nucleotides, compatible with the previous reports. The recombinant plasmid pET-22b(+)/AB was successfully constructed. DNA sequencing showed one open reading frame with the length of 588 bp. It encoded seven conservative regions that showed good antigenicity and hydrophobicity by Parker and Welling method. Furthermore, INTERNET EXPASY, NNPREDICT and ISREC predicted that it was a porin-like structure consisting of beta-pleated sheets that were embedded in the outer membrane. BLAST analyzed 836 767 protein sequences and found that the similar sequences were all belonging to H pylori OMP sequences. SDS-PAGE and scan analysis showed that the molecular weight of AB was 22.5 ku and recombinant protein amounted to 29% of the total bacterial protein, among which dissolved expression amounted to 21.9% of sonicated supernatant. The rAB purity amounted to 96% through affinity chromatography. Western blot analysis of rAB confirmed that it could be specially recognized by serum form rabbit immunized with AlpA and H pylori infected. CONCLUSION: Adhesin AlpA recombinant protein may be a potential vaccine for control and treatment of H pylori infection.
机译：目的：构建表达幽门螺杆菌黏附素AlpA的重组菌株，研究黏附素AlpA的免疫原性。方法：通过PCR技术从幽门螺杆菌的染色体DNA中扩增出基因Ab，并对其生物学信息进行分析，并使用T4 DNA将其插入表达载体pET-22b（+）的Nco I和Not I限制性片段中。连接酶。使用氨苄青霉素抗性进行选择，将所得质粒pET-AlpA转化到感受态大肠杆菌BL21（DE3）细胞中。将重组菌株在100 mL / mL氨苄青霉素的5 mL LB中于37度孵育过夜。通过蛋白质印迹法分析BL21（DE3）pET-22b（+）/ AlpA的超声处理，以检测AlpA的免疫原性。结果：以幽门螺杆菌悉尼菌株（SS1）的染色体DNA为模板，通过PCR扩增了AlpA蛋白编码基因。结果表明，经PCR扩增的AlpA DNA片段约有1 500个核苷酸，与以前的报道相符。成功构建了重组质粒pET-22b（+）/ AB。 DNA测序显示一个开放阅读框，长度为588 bp。它通过Parker和Welling方法编码了七个显示出良好抗原性和疏水性的保守区域。此外，INTERNET EXPASY，NNPREDICT和ISREC预测这是一种类似孔蛋白的结构，由嵌入在外膜中的β折叠薄片组成。 BLAST分析了836767个蛋白质序列，发现相似的序列全部属于幽门螺杆菌OMP序列。 SDS-PAGE和扫描分析表明，AB的分子量为22.5 ku，重组蛋白占细菌总蛋白的29％，其中溶解表达占超声上清液的21.9％。通过亲和层析，rAB纯度为96％。对rAB的Western印迹分析证实，它可以被AlpA和幽门螺杆菌感染的兔免疫血清形式特异性识别。结论：粘附素AlpA重组蛋白可能是控制和治疗幽门螺杆菌感染的潜在疫苗。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号