...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Journal of Pure & Applied Microbiology >Cloning, Expression and Purification of HpaA Recombinant Protein of Helicobacter pylori as a Vaccine Candidate
【24h】

Cloning, Expression and Purification of HpaA Recombinant Protein of Helicobacter pylori as a Vaccine Candidate

机译：幽门螺杆菌HpaA重组蛋白的克隆，表达及纯化

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

Helicobacter pylori is prevailing in gastric niches and chronic disease with this pathogen can cause increased risk of many diseases such as gastritis, duodenal ulcer, dysplasia, neoplasia, gastric B cell lymphoma. Emergence of antibiotic resistance makes to recommended different treatments, and prevention methods have offered against infection. Today, among these methods as a result of immunization both in prevention and treatment of the infection has been confirmed and great efforts for vaccine design have been made. This study aimed to evaluate the cloning, expression and purification of HpaA recombinant protein of H. pylori as a vaccine candidate. Bioinformatics designing for desired gene were performed by use of suitable softwares and the construct was ordered to shine company (China), after receiving construct, cloned in pET21b vector, and this process was confirmed by polymerase chain reaction (PCR), digestion and sequencing techniques. Then, it was successfully cloned in the Escherichia coli B1.-21 as expression host and the protein expressed. Expression of protein was verified using SDS page and western blot, for purification of protein, the Ni-NTA column was used. Dialysis for removal of imidazole was applied. Finally, protein concentration was determined by Bicinchoninic Acid Protein assay Kit (Parstoos). In present study, HpaA recombinant protein with size 29 kDa was expressed and purified, successfully. In our study construct HpaA was synthesized, but complementary studies are necessary to assess the immunological characteristics of this construct as an appropriate and efficient vaccine candidate against H. pylori.
机译：幽门螺杆菌在胃ni中盛行，这种病原体的慢性疾病可导致许多疾病的风险增加，例如胃炎，十二指肠溃疡，发育异常，瘤形成，胃B细胞淋巴瘤。抗生素耐药性的出现导致推荐了不同的治疗方法，并且提供了预防感染的方法。今天，在这些方法中，作为预防和治疗感染的免疫方法的结果已经得到证实，并且已经为疫苗设计做出了巨大的努力。本研究旨在评估幽门螺杆菌HpaA重组蛋白作为候选疫苗的克隆，表达和纯化。使用合适的软件进行所需基因的生物信息学设计，并在接到构建体后命令该构建体向shine company（中国）克隆到pET21b载体中，并通过聚合酶链反应（PCR），消化和测序技术证实了这一过程。然后，将其成功克隆到大肠杆菌B1.-21中作为表达宿主并表达了蛋白质。使用SDS页和Western blot验证蛋白质的表达，使用Ni-NTA柱纯化​​蛋白质。进行透析以去除咪唑。最后，通过比辛可宁酸蛋白质测定试剂盒（Parstoos）测定蛋白质浓度。在本研究中，成功​​表达并纯化了大小为29 kDa的HpaA重组蛋白。在我们的研究中，合成了HpaA构建体，但需要进行补充研究以评估该构建体作为针对幽门螺杆菌的合适而有效的疫苗候选者的免疫学特征。

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