Evaluation of the eukaryotic expression of mtb32C-hbha fusion gene of Mycobacterium tuberculosis in Hepatocarcinoma cell line
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 tw...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Tehran University of Medical Sciences
2016-05-01
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| Series: | Iranian Journal of Microbiology |
| Subjects: | |
| Online Access: | https://ijm.tums.ac.ir/index.php/ijm/article/view/118 |
| Summary: | 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.
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| ISSN: | 2008-3289 2008-4447 |