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首页> 外文期刊>Iranian Journal of Microbiology >Evaluation of the eukaryotic expression of mtb32C-hbha fusion gene of Mycobacterium tuberculosis in Hepatocarcinoma cell line
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Evaluation of the eukaryotic expression of mtb32C-hbha fusion gene of Mycobacterium tuberculosis in Hepatocarcinoma cell line

机译：结核分枝杆菌mtb32C-hbha融合基因在肝癌细胞系中的真核表达评估

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Background and Objectives: HBHA and Mtb32C have been isolated from culture supernatants of Mycobacterium tu- berculosis (M. tuberculosis) and Mycobacterium bovis (M. bovis) and their immunogenicity previously studies have been confirmed. In this study, capability of constructed vector containing two mycobacterial immunodaminant antigens (Mt- b32C-HBHA), in producing new chimeric protein under the in-vitro condition was examined.Materials and Methods: In present study Huh7.5 cells was transfected with Mtb32C-HBHA –pCDNA3.1+ recombinant vector using the calcium phosphate method and expression of chimeric protein was assessed by RT-PCR and Western blot methods.Results: Results of RT-PCR and Western blot showed expression of 35.5 KD recombinant protein (Mtb32C-HBHA) in thiscell line.Conclusion: The constructed vector can produce two highly immunogenic antigens that fusion of them to gather makes chi- meric antigen with new traits. Other attempts are needed to evaluate specific properties of this new antigen such as molecular conformation modeling and immunologic characteristics in future studies.
机译：背景与目的：已从结核分枝杆菌（结核分枝杆菌）和牛分枝杆菌（牛分枝杆菌）的培养上清液中分离了HBHA和Mtb32C，并已确认了它们的免疫原性。在这项研究中，研究了含有两种分枝杆菌免疫损伤抗原（Mt-b32C-HBHA）的载体在体外条件下产生新嵌合蛋白的能力。材料与方法：在本研究中，将Huh7.5细胞用Mtb32C转染RT-PCR和Western blot方法检测磷酸钙-HBHA –pCDNA3.1 +重组载体的表达及嵌合蛋白的表达。结果：RT-PCR和Western blot结果显示35.5 KD重组蛋白（Mtb32C-结论：构建的载体可以产生两种高度免疫原性的抗原，它们融合在一起会产生具有新特性的嵌合抗原。还需要其他尝试来评估这种新抗原的特定特性，例如在未来的研究中进行分子构象建模和免疫学特征。

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