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首页> 外文期刊>Iranian journal of public health. >High-Level Expression of Immunogenic Recombinant Plasmodium vivax Merozoite Surface Protein (Pvmsp-142 kDa) in pGEX 6P1 Vector
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High-Level Expression of Immunogenic Recombinant Plasmodium vivax Merozoite Surface Protein (Pvmsp-142 kDa) in pGEX 6P1 Vector

机译：免疫原性重组间日疟原虫裂殖子表面蛋白（Pvmsp-1 42 kDa）在pGEX 6P1载体中的高表达

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BackgroundDetection of Plasmodium vivax specific antibodies with serological tests could be a valuable tool for epidemiological researches. Whereas P. vivax cannot be simply obtained in vitro, serological tests using total or semi-purified antigens are infrequently used. Given this restriction, the present study investigated whether recombinant P. vivax merozoite surface protein 1 (PvMSP-1 42 kDa) could be useful in detection of antibodies from the serums of a P. vivax infected person using serological tests.MethodsParasite DNA was extracted from blood sample of an Iranian P. vivax-infected patient. The region of PvMSP-142 kDa was amplified by PCR then cloned into pTZ57R/T vector and sequenced. The insert was sub cloned into pGEX 6P1 expression vector. Afterwards, it was transformed into E. coli BL21 and cultured massively. Sub cloning of gene was confirmed by PCR and enzyme digestion and sequencing finally. Production of recombinant protein was confirmed by SDS-PAGE. Western blot was performed by human sera to appraisal binding ability to the IgG antibodies of P. vivax infected patients. Recombinant protein was purified and estimated by Bradford assay.Results:The specialty values of the Western blot determined with 10 sera from naturally infected individuals, 10 sera from healthy individuals and 7 sera from individuals with other infectious diseases.ConclusionFor the Iranian population, using a Western blot assay for MSP-142 recombinant protein can be used as the foundation for promotion of serological assay for the detection of P. vivax malaria such as ELISA.
机译：背景血清学检测检测间日疟原虫特异性抗体可能是流行病学研究的重要工具。尽管间日疟原虫不能简单地在体外获得，但很少使用使用总抗原或半纯化抗原的血清学测试。在这种限制下，本研究调查了重组间日疟原虫裂殖子表面蛋白1（PvMSP-1 42 kDa）是否可用于通过血清学检测从间日疟原虫感染者的血清中检测抗体的方法。一名伊朗间日疟原虫感染患者的血液样本。通过PCR扩增PvMSP-142kDa的区域，然后克隆到pTZ57R / T载体中并测序。将插入物亚克隆到pGEX 6P1表达载体中。之后，将其转化到大肠杆菌BL21中并进行大规模培养。通过PCR，酶切和测序最终确定了基因的亚克隆。通过SDS-PAGE确认重组蛋白的产生。通过人血清进行蛋白质印迹，以评估与间日疟原虫感染患者的IgG抗体的结合能力。结果：用自然感染个体的10份血清，健康个体的10份血清和其他传染病个体的7份血清测定了Western blot的特异性值。结论对于伊朗人群，使用MSP-142重组蛋白的蛋白质印迹法可作为促进血清学检测（例如ELISA）的血清学检测的基础。

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