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首页> 外文期刊>Asian Journal of Pharmaceutical and Clinical Research >CLONINIG AND EXPRESSION OF BAB-PATHOGENICITY ISLAND ANTIGENS FOR THE PRODUCTION OF VACCINE AGAINST HELICOBACTER PYLORI, THE RISK FACTOR FOR GASTRIC CANCER
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CLONINIG AND EXPRESSION OF BAB-PATHOGENICITY ISLAND ANTIGENS FOR THE PRODUCTION OF VACCINE AGAINST HELICOBACTER PYLORI, THE RISK FACTOR FOR GASTRIC CANCER

机译:BAB-致病性岛抗原的克隆和表达,用于生产针对幽门螺杆菌的疫苗,幽门螺杆菌是胃癌的危险因素

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摘要

BabA2, one of the two allele of BabA gene and a member protein of Helicobacter pylori, plays a vital role in assisting bacterial colonization in the stomach. However, its association with H pylori -related gastroduodenal diseases still remains unclear. In the present study, babA2 gene from Helicobacter pylori was amplified using specific primers and then cloned in pTZ57R/T and transformed into DH5-α cells successfully. Transformation was confirmed with plasmid extraction and followed by restriction digestion. IPTG was used as an inducer for the expression of babA2 protein and the protein was successfully isolated and quantified. The quantified protein was subjected to SDS PAGE to evaluate the expression of that protein. The sequence analysis have shown 99% “perfect” match with sequences of their corresponding gene (babA2) from GenBank as determined by using BLAST (version 2.7). Inserted babA2 gene was expressed significantly in the prokaryotic expression system, and specific strip at ~ 75 KDa was demonstrated in SDS-PAGE. Further study is needed to substantiate that the expressed protein will act as an antigen for the humoral immunity against the Helicobacter pylori.
机译:BabA2是BabA基因的两个等位基因之一,是幽门螺杆菌的成员蛋白,在协助细菌在胃中定殖方面起着至关重要的作用。然而,其与幽门螺杆菌相关的胃十二指肠疾病的关系仍不清楚。在本研究中,使用特异性引物扩增了幽门螺杆菌的babA2基因,并将其克隆到pTZ57R / T中并成功转化入DH5-α细胞。通过质粒提取确认转化,然后进行限制性消化。 IPTG用作babA2蛋白表达的诱导剂,该蛋白已成功分离和定量。将定量的蛋白质进行SDS PAGE以评估该蛋白质的表达。序列分析显示,与使用BLAST(2.7版)确定的来自GenBank的相应基因(babA2)的序列有99%的“完美”匹配。插入的babA2基因在原核表达系统中表达明显,并在SDS-PAGE中显示〜75 KDa的特异性条带。需要进一步的研究来证实表达的蛋白质将作为针对幽门螺杆菌的体液免疫的抗原。

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