[Home ] [Archive]   [ فارسی ]  
:: Main :: About :: Current Issue :: Archive :: Search :: Submit :: Contact ::
Main Menu
Home::
Journal Information::
Articles archive::
Publication Ethics::
Peer Review Process::
Indexing Databases::
For Authors::
For Reviewers::
Subscription::
Contact us::
Site Facilities::
::
Google Scholar Metrics

Citation Indices from GS

AllSince 2019
Citations62733583
h-index2719
i10-index18478

..
Search in website

Advanced Search
..
Receive site information
Enter your Email in the following box to receive the site news and information.
..
Registered in

AWT IMAGE

AWT IMAGE

..
:: Volume 23, Issue 7 (2-2016) ::
Journal of Ilam University of Medical Sciences 2016, 23(7): 18-27 Back to browse issues page
Production of Recombinant Construct by Cloning of Protective Antigen Domain 4 Gene and Fusion of it with Lethal Factor Domain 1 Gene of Bacillus anthracis in E.coli
Mohammad Najarasl1 , Mohammad Sadegh Hashemzadeh2 , Hosein Honari * 3, Jafar Mousavy1 , Firouz Ebrahimi1 , Hosein Pourhakkak1
1- Comprehensive University of Imam Hossein, Tehran, Iran.
2- Baqiyatallah University of Medical Sciences, Tehran, Iran.
3- Comprehensive University of Imam Hossein, Tehran, Iran. , msh.biotechnology@gmail.com
Abstract:   (6926 Views)

Introduction: Anthrax is a zoonotic disease. Bacterium Bacillus anthracis is the causative agent of the fatal disease. At present, the protective antigen (PA) is used as an effective vaccine against anthrax. Domain 4 of this antigen together with domain 1 of lethal factor (LF) are the potent immunogens of this bacteria and are as suitable candidates of vaccine against it. Our aim in this study is the cloning of protective antigen domain 4 (PAD4) genes and fusion of it with lethal factor domain 1 (LFD1) gene of the bacteria to evaluate their capability in protective immunity induction.

Materials & methods: In this experimental study, we used a recombinant pGEM-T easy vector containing LFD1 gene. Then PAD4 gene was amplified and isolated by PCR and cloned into another pGEM-T easy vector, separately. After that, ligation of PAD4 gene and LFD1 gene was done in mentioned vector with determination of LFD1 orientation and by PstI/XbaI restriction sites. The recombinant construct, resulted from these genes was sub-cloned into pET28a expression vector using BamHI/ XhoI restriction enzymes and after determination of genes orientation, the expression host BL21 was transformed by this recombinant vector.

Findings: First cloning and fusion of PAD4 and LFD1 gene fragments were successfully carried out in pGEM-T easy vector and after the confirmation of mentioned process by both of enzymatic digestion and PCR methods, the result recombinant construct was sub-cloned into pET28a.

Discussion & Conclusions: Since PAD4 and LFD1 are immunogenic regions, expression of the recombinant construct resulted from these ligated genes can be proposed as proper candidate of anthrax vaccine for induction of protective immunity.

Keywords: Bacillus anthracis, Protective antigen (PA), Lethal factor (LF), Fusion, Vaccine candidate
Full-Text [PDF 699 kb]   (2294 Downloads)    
Type of Study: Research | Subject: Bacteriology
Received: 2015/05/14 | Accepted: 2015/06/30 | Published: 2016/02/14
Send email to the article author

Add your comments about this article
Your username or Email:

CAPTCHA


XML   Persian Abstract   Print


Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:

Najarasl M, Hashemzadeh M S, Honari H, Mousavy J, Ebrahimi F, Pourhakkak H. Production of Recombinant Construct by Cloning of Protective Antigen Domain 4 Gene and Fusion of it with Lethal Factor Domain 1 Gene of Bacillus anthracis in E.coli. J. Ilam Uni. Med. Sci. 2016; 23 (7) :18-27
URL: http://sjimu.medilam.ac.ir/article-1-2702-en.html


Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Volume 23, Issue 7 (2-2016) Back to browse issues page
مجله دانشگاه علوم پزشکی ایلام Journal of Ilam University of Medical Sciences
Persian site map - English site map - Created in 0.15 seconds with 41 queries by YEKTAWEB 4643