[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
Citations62753585
h-index2719
i10-index18578

..
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 27, Issue 5 (12-2019) ::
Journal of Ilam University of Medical Sciences 2019, 27(5): 24-36 Back to browse issues page
Subcloning and Expression of the Chimeric Antigen C-CFTX1-STxB of the Jellyfish Venom and its Antigenicity Assessment in Syrian Mice
Hossein Honari * 1, Seyed Mojtaba Aghaie2 , Mehdi Hosseinzade2
1- Biological Research Center, Faculty of Basic Sciences, Imam Hossein Comprehensive University, Tehran, Iran , honari.hosein@gmail.com
2- Biological Research Center, Faculty of Basic Sciences, Imam Hossein Comprehensive University, Tehran, Iran
Abstract:   (2891 Views)
Introduction: Box jellyfish stings are painful and may be life-threatening. The venom of Chironex fleckeri contains a variety of bioactive proteins as well as two of the most abundant proteins, namely CfTX-1 and CfTX-2 which cannot be isolated easily using electrophoresis or chromatography techniques. Recombinant expression technology may offer an alternative to the isolation of native C.fleckeri venom protein. This study aimed at expressing C-CfTX1-STxB protein in Escherichia coli and assessing its antigenicity in Syrian mice.
 
Materials & Methods: Synthesis of the artificial CfTX1complete gene was prepared in plasmid pUC57. The C-cftx1 was cloned using a polymerase chain reaction (PCR) and subcloned with BamHI and SalI restriction enzyme sites in pET28a-stxB expression vector and transformed into E.coli. Gene expression was artificially induced by Isopropyl β- d-1-thiogalactopyranoside. After the purification of the protein and its injection into the Syrian mice, the amount of produced antibody was measured in the serum. The rats were also challenged by the venom of the jellyfish (i.e., Rhopilema nomadic).
 
Findings: In this experimental study, the C-CfTX1-STxB gene was cloned in the expression vector pET28a (+), sequenced by PCR, and analyzed by enzymatic analysis. Moreover, the produced recombinant protein was confirmed by Western blotting. The produced antibody in the serum was quantified using an enzyme-linked immunosorbent assay.
 
Discussion & Conclusions: After 60 days, the immunized mice tolerated 50x LD50 of jellyfish venom. Considering the ineffectiveness of cardiotoxicity and neurotoxicity of the recombinant protein, this produced protein can be suggested as a jellyfish venom vaccine candidate in Syrian mice or at a later stage of a clinical trial in humans.
Keywords: Antigenicity, C-CFTX1-STxB chimeric antigen, Chironex fleckeri, Jellyfish venom
Full-Text [PDF 930 kb]   (859 Downloads)    
Type of Study: Research | Subject: biology
Received: 2018/12/23 | Accepted: 2019/04/8 | Published: 2019/12/31
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:

Honari H, Aghaie S M, Hosseinzade M. Subcloning and Expression of the Chimeric Antigen C-CFTX1-STxB of the Jellyfish Venom and its Antigenicity Assessment in Syrian Mice. J. Ilam Uni. Med. Sci. 2019; 27 (5) :24-36
URL: http://sjimu.medilam.ac.ir/article-1-5331-en.html


Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Volume 27, Issue 5 (12-2019) 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