Volume 48, Issue 2 (8-2024)
Research in Medicine 2024, 48(2): 27-36 |
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Ethics code: IR.IAU.KERMAN.REC.1401.078
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Sadeghi S, Khandan Dezfully N, Amini K. Molecular Study and Cloning of the dt Gene of Corynebacterium in Susceptible Cells in Order to Prepare a Vaccine. Research in Medicine 2024; 48 (2) :27-36
URL: http://pejouhesh.sbmu.ac.ir/article-1-3299-en.html
URL: http://pejouhesh.sbmu.ac.ir/article-1-3299-en.html
Department of Microbiology, Sirjan Branch, Islamic Azad University, Sirjan, Iran. , nooshinkhandan22@gmail.com
Abstract: (378 Views)
Background and Aim: Corynebacterium diphtheriae is an aerobic, gram-positive, non- sporulating bacterium. Diphtheria toxin (dt) is a highly toxic protein and produced as a result of bacterial infection with a special corynephage that carries tox. The aim of the present study was to clone the dt gene of Corynebacterium in susceptible E.coli XL1blue cell in order to produce a recombinant protein and use this protein in subsequent research in 2022 at Pasargård Research Institute.
Methods: In this Cross- sectional study, a DNA sample of Corynebacterium diphtheria vaccinal strain PW8 Sub- strain 2000CN was prepared from Razi Institute in Tehran. To identify the dt gene, the DNA bacterial strain was amplified using specific primers in the polymerase chain reaction (PCR) method. Using the TA- cloning kit, the dt gene was cloned in the susceptible E.coli XL1blue cell. The phylogeny tree for the bacterial strain was performed by DNA sequencing of the 16 srRNA gene region using clustalX and Mega5 software.
Results: The DNA of Corynebacterium diphtheria carried the dt gene. TA- cloning of the dt gene was confirmed by the Real- Time PCR method, DNA sequencing of the PCR product, the presence of white- blue colonies, and amplification with an M13 primer. The result of DNA sequencing of the 16 srRNA gene region was confirmed as Corynebacterium diphtheria.
Conclusion: In this study, the dt gene was cloned in the E.coli XL1blue vector and then confirmed, and it can be considered as a powerful antigen for the preparation of the recombinant protein tool, as well as for stimulating the immune system. This gene can be used in future studies.
Methods: In this Cross- sectional study, a DNA sample of Corynebacterium diphtheria vaccinal strain PW8 Sub- strain 2000CN was prepared from Razi Institute in Tehran. To identify the dt gene, the DNA bacterial strain was amplified using specific primers in the polymerase chain reaction (PCR) method. Using the TA- cloning kit, the dt gene was cloned in the susceptible E.coli XL1blue cell. The phylogeny tree for the bacterial strain was performed by DNA sequencing of the 16 srRNA gene region using clustalX and Mega5 software.
Results: The DNA of Corynebacterium diphtheria carried the dt gene. TA- cloning of the dt gene was confirmed by the Real- Time PCR method, DNA sequencing of the PCR product, the presence of white- blue colonies, and amplification with an M13 primer. The result of DNA sequencing of the 16 srRNA gene region was confirmed as Corynebacterium diphtheria.
Conclusion: In this study, the dt gene was cloned in the E.coli XL1blue vector and then confirmed, and it can be considered as a powerful antigen for the preparation of the recombinant protein tool, as well as for stimulating the immune system. This gene can be used in future studies.
Type of Study: Original |
Subject:
Microbiology
Received: 2023/06/30 | Accepted: 2024/06/30 | Published: 2024/09/16
Received: 2023/06/30 | Accepted: 2024/06/30 | Published: 2024/09/16
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