year 4, Issue 3 (Fall 2010)
Iran J Med Microbiol 2010, 4(3): 36-44 |
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Ghaderi H, Rezaie S, Noorbakhsh F, Vafaie M, Motavaze K, Emamy M et al . Cloning of G subunit’s of V-ATPase gene from Trychophyton rubrum for use the proteins. Iran J Med Microbiol 2010; 4 (3) :36-44
URL: http://ijmm.ir/article-1-38-en.html
URL: http://ijmm.ir/article-1-38-en.html
Hajarasadat Ghaderi 
1, Sasan Rezaie2 , Fatemeh Noorbakhsh3 , Mahtab Vafaie1 , Kamiyar Motavaze1 , Masoud Emamy1 , Mohammadreza Safari
1, Sasan Rezaie2 , Fatemeh Noorbakhsh3 , Mahtab Vafaie1 , Kamiyar Motavaze1 , Masoud Emamy1 , Mohammadreza Safari
1- Department of Microbiology, Islamic Azad university, North Tehran branche, Tehran, Iran
2- Department of Mycology and Parazitology, Molecular biology section, Faculty of Health, Tehran university of medical sciences, Tehran, Iran
3- Department of Microbiology, Islamic Azad university, Varamin-Pishva branche, Varamin, Iran
2- Department of Mycology and Parazitology, Molecular biology section, Faculty of Health, Tehran university of medical sciences, Tehran, Iran
3- Department of Microbiology, Islamic Azad university, Varamin-Pishva branche, Varamin, Iran
Abstract: (14865 Views)
Background & objective: Trichophyton rubrum is the most common etiologic agent of chronic dermatophytosis of body in the world. Therapy of this disease is not complete and the disease may relapse after stopping medicine consumption. Recently it is suggested that V-ATPase may work as a target for antifungal drugs development. In this study we cloned G subunit of V-ATPase gene from T.rubrum into E.coli to produce the relevant protein in high density, save it and then study it.
Material and methods: DNA and RNA was extracted from Trichophyton rubrum. Then, the gene was replicated using PCR & RT-PCR. We inserted cDNA in pTG19-T and cloned it in E.coli white using hit shock method. Then PCR was done with the same protocol.
Results: DNA has 750 bp and cDNA has 700 bp. When E. coli was cultured on Luria agar we found both blue and white colonies and by doing PCR it was shown that the 5 colony of E.coli had this gene.
Conclusion: It seems the gene V-ATPase has intron. It is also suggests that the white colonies have probably the gene. The product of latest PCR indicated that the cloning was done successfully.
Material and methods: DNA and RNA was extracted from Trichophyton rubrum. Then, the gene was replicated using PCR & RT-PCR. We inserted cDNA in pTG19-T and cloned it in E.coli white using hit shock method. Then PCR was done with the same protocol.
Results: DNA has 750 bp and cDNA has 700 bp. When E. coli was cultured on Luria agar we found both blue and white colonies and by doing PCR it was shown that the 5 colony of E.coli had this gene.
Conclusion: It seems the gene V-ATPase has intron. It is also suggests that the white colonies have probably the gene. The product of latest PCR indicated that the cloning was done successfully.
Type of Study: Original Research Article |
Subject:
Medical Bacteriology
Received: 2013/10/28 | Accepted: 2013/11/10 | ePublished: 2013/11/10
Received: 2013/10/28 | Accepted: 2013/11/10 | ePublished: 2013/11/10
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