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首页> 外文期刊>Molecular Biology Reports >Expression of UreB and HspA of Helicobacter pylori in silkworm pupae and identification of its immunogenicity
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Expression of UreB and HspA of Helicobacter pylori in silkworm pupae and identification of its immunogenicity

机译：幽门螺杆菌UreB和HspA在家蚕p中的表达及免疫原性鉴定

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摘要

For mass production of urease B subunit (UreB) and heat shock protein A subunit (HspA) of Helicobacter pylori with Bombyx mori nuclear polyhedrosis virus (BmNPV) baculovirus expression system (BES) and to determine whether they could be used as an oral vaccine against H. pylori, besides, to determine the time course of expressed recombinant protein and the optimum acquisition time directly through green fluorescence, HspA and enhanced green fluorescence protein (EGFP) genes were cloned into vector pFastBacDual to form donor vector pFastBacDual-(EGFP) (HspA), UreB gene was cloned into vector pFastBacDual to form donor vector pFastBacDual-UreB,then they were transformed into E. coli BmDH10Bac to obtain the recombinant Bacmid-(EGFP) (HspA) and Bacmid-UreB respectively. They were used to transfect BmN cells and generated the recombinant baculovirus BmNPV-(EGFP) (HspA) and BmNPV-UreB. Using these recombinant baculovirus BmNPV-(EGFP) (HspA) and BmNPV-UreB inoculated the silkworm pupae, a recombinant HspA and UreB protein were expressed in silkworm pupae, which were around 13 and 62 kDa in sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analysis. After oral immunization of mice, serum specific IgG antibodies against HspA and UreB in vaccine group were much higher than that in mock and native silkworm powder control groups. The results indicated that the expressed recombinant HspA and UreB in silkworm pupae would possess good immunogenicity. In addition, when EGFP and HspA proteins were expressed, a direct correlation between the increase in intensity of fluorescence and HspA concentration.
机译：用于批量生产带有家蚕核多角体病毒（BmNPV）杆状病毒表达系统（BES）的幽门螺杆菌的脲酶B亚基（UreB）和热休克蛋白A亚基（HspA），并确定它们是否可以用作针对此外，为了确定幽门螺杆菌的表达时间，并直接通过绿色荧光确定最佳的获取时间，将HspA和增强型绿色荧光蛋白（EGFP）基因克隆到载体pFastBacDual中以形成供体载体pFastBacDual-（EGFP）（ HspA），将UreB基因克隆到载体pFastBacDual中形成供体载体pFastBacDual-UreB，然后将其转化到大肠杆菌BmDH10Bac中，分别得到重组Bacmid-（EGFP）（HspA）和Bacmid-UreB。它们被用于转染BmN细胞并产生重组杆状病毒BmNPV-（EGFP）（HspA）和BmNPV-UreB。使用这些重组杆状病毒BmNPV-（EGFP）（HspA）和BmNPV-UreB接种家蚕p，在家蚕p中表达了重组HspA和UreB蛋白，在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS）中分别为13和62 kDa。 -PAGE）和蛋白质印迹分析。小鼠口服免疫后，疫苗组中针对HspA和UreB的血清特异性IgG抗体要远远高于模拟和天然蚕粉对照组。结果表明，在家蚕p中表达的重组HspA和UreB具有良好的免疫原性。此外，当表达EGFP和HspA蛋白时，荧光强度的增加与HspA浓度之间存在直接关系。

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