Background: IL-2 is a cytokine that plays an active role in the activation and maintenance of both acquired and innate immune defenses. It is also capable of improving the protective immune responses that are generated by conventional vaccines against avian pathogens in the poultry industry when used as an adjuvant.
Objectives: The aim of this study was to extract and sequence Iranian chicken IL-2.
Materials and Methods: In this study, genomic DNA was extracted from Iranian chickens. Total RNA was isolated by culturing harvested splenocytesand lysing them with Trizol reagent per the manufacturer's instructions. mRNA was isolated and converted to cDNA using reverse-transcriptase (RT) and specific designed primers. Then, the PCR product was ligated into the PTZ57R/T plasmid (TA-cloning) and transformed into competent Top10 E. coli cells.
Results: A unique 668-bp band was obtained after RT-PCR. Restriction enzyme digestion and colony PCR analysis and direct sequencing confirmed the existence of the desired gene in transformants.
Conclusions: For first time in Iran, the chIL-2 gene was successfully extracted, cloned, and transferred into E. coli. The efficacy of recombinant or DNA vaccines can be modulated by codelivery of this cytokine gene.


Implication for health policy/practice/research/medical education:
The ability of chIL-2 to enhance T-cell activation and proliferation suggests that it augments the immune response when used as an adjuvant.
Please cite this paper as:
Amini H, Hoseini SD, Nowroozi J, Shahbazzadeh D. Cloning and Sequencing of Iranian Chicken Interleukine-2 Gene. Jundishapur J Microbiol.2012;5(3): 502-6. DOI: 10.5812/jjm.3636

Copyright ©2012 Kowsar Corp. All rights reserved.


"/> Background: IL-2 is a cytokine that plays an active role in the activation and maintenance of both acquired and innate immune defenses. It is also capable of improving the protective immune responses that are generated by conventional vaccines against avian pathogens in the poultry industry when used as an adjuvant.
Objectives: The aim of this study was to extract and sequence Iranian chicken IL-2.
Materials and Methods: In this study, genomic DNA was extracted from Iranian chickens. Total RNA was isolated by culturing harvested splenocytesand lysing them with Trizol reagent per the manufacturer's instructions. mRNA was isolated and converted to cDNA using reverse-transcriptase (RT) and specific designed primers. Then, the PCR product was ligated into the PTZ57R/T plasmid (TA-cloning) and transformed into competent Top10 E. coli cells.
Results: A unique 668-bp band was obtained after RT-PCR. Restriction enzyme digestion and colony PCR analysis and direct sequencing confirmed the existence of the desired gene in transformants.
Conclusions: For first time in Iran, the chIL-2 gene was successfully extracted, cloned, and transferred into E. coli. The efficacy of recombinant or DNA vaccines can be modulated by codelivery of this cytokine gene.


Implication for health policy/practice/research/medical education:
The ability of chIL-2 to enhance T-cell activation and proliferation suggests that it augments the immune response when used as an adjuvant.
Please cite this paper as:
Amini H, Hoseini SD, Nowroozi J, Shahbazzadeh D. Cloning and Sequencing of Iranian Chicken Interleukine-2 Gene. Jundishapur J Microbiol.2012;5(3): 502-6. DOI: 10.5812/jjm.3636

Copyright ©2012 Kowsar Corp. All rights reserved.


"/>

Cloning and Sequencing of Iranian Chicken Interleukine-2 Gene

AUTHORS

Hamideh Amini 1 , * , Seyed Davood Hoseini 2 , Jamileh Nowroozi 3 , Delavar Shahbazzadeh 3

1 Department of Microbiology, Faculty of Biology, Islamic Azad University, Tehran Branch, [email protected], IR Iran

2 Department of Cellular And Molecular Biology, Razi vaccination and serum research institute, IR Iran

3 Department of Microbiology, Faculty of Biology, Islamic Azad University, Tehran Branch, IR Iran

How to Cite: Amini H, Hoseini S, Nowroozi J, Shahbazzadeh D. Cloning and Sequencing of Iranian Chicken Interleukine-2 Gene, Jundishapur J Microbiol. Online ahead of Print ; 5(3):502-506. doi: 10.5812/jjm.3636.

ARTICLE INFORMATION

Jundishapur Journal of Microbiology: 5 (3); 502-506
Article Type: Research Article
Received: April 10, 2011
Accepted: December 1, 2011
Crossmark
Crossmark
CHECKING
READ FULL TEXT

Abstract

Background: IL-2 is a cytokine that plays an active role in the activation and maintenance of both acquired and innate immune defenses. It is also capable of improving the protective immune responses that are generated by conventional vaccines against avian pathogens in the poultry industry when used as an adjuvant.
Objectives: The aim of this study was to extract and sequence Iranian chicken IL-2.
Materials and Methods: In this study, genomic DNA was extracted from Iranian chickens. Total RNA was isolated by culturing harvested splenocytesand lysing them with Trizol reagent per the manufacturer's instructions. mRNA was isolated and converted to cDNA using reverse-transcriptase (RT) and specific designed primers. Then, the PCR product was ligated into the PTZ57R/T plasmid (TA-cloning) and transformed into competent Top10 E. coli cells.
Results: A unique 668-bp band was obtained after RT-PCR. Restriction enzyme digestion and colony PCR analysis and direct sequencing confirmed the existence of the desired gene in transformants.
Conclusions: For first time in Iran, the chIL-2 gene was successfully extracted, cloned, and transferred into E. coli. The efficacy of recombinant or DNA vaccines can be modulated by codelivery of this cytokine gene.


Implication for health policy/practice/research/medical education:
The ability of chIL-2 to enhance T-cell activation and proliferation suggests that it augments the immune response when used as an adjuvant.
Please cite this paper as:
Amini H, Hoseini SD, Nowroozi J, Shahbazzadeh D. Cloning and Sequencing of Iranian Chicken Interleukine-2 Gene. Jundishapur J Microbiol.2012;5(3): 502-6. DOI: 10.5812/jjm.3636

Copyright ©2012 Kowsar Corp. All rights reserved.


Full Text

Full text is available in PDF

© 0, Author(s). This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/) which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited.
COMMENTS

LEAVE A COMMENT HERE: