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Molecular Cloning, Expression, and In Silico Structural Analysis of Guinea Pig IL-17

Dirisala, Vijaya R. and Jeevan, Amminikutty and Ramasamy, Suresh K. and McMurray, David N. (2013) Molecular Cloning, Expression, and In Silico Structural Analysis of Guinea Pig IL-17. Molecular Biotechnology, 55 (3). pp. 277-287. ISSN 1073-6085. https://resolver.caltech.edu/CaltechAUTHORS:20140110-140009344

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Abstract

Interleukin-17A (IL-17A) is a potent proinflammatory cytokine and the signature cytokine of Th17 cells, a subset which is involved in cytokine and chemokine production, neutrophil recruitment, promotion of T cell priming, and antibody production. IL-17 may play an important role in tuberculosis and other infectious diseases. In preparation for investigating its role in the highly relevant guinea pig model of pulmonary tuberculosis, we cloned guinea pig IL-17A for the first time. The complete coding sequence of the guinea pig IL-17A gene (477 nucleotides; 159 amino acids) was subcloned into a prokaryotic expression vector (pET-30a) resulting in the expression of a 17 kDa recombinant guinea pig IL-17A protein which was confirmed by mass spectrometry analysis. Homology modeling of guinea pig IL-17A revealed that the three-dimensional structure resembles that of human IL-17A. The secondary structure predicted for this protein showed the presence of one extra helix in the N-terminal region. The expression profile of IL-17A was analyzed quantitatively in spleen, lymph node, and lung cells from BCG-vaccinated guinea pigs by real-time PCR. The guinea pig IL-17A cDNA and its recombinant protein will serve as valuable tools for molecular and immunological studies in the guinea pig model of pulmonary TB and other human diseases.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1007/s12033-013-9679-zDOIArticle
http://link.springer.com/article/10.1007/s12033-013-9679-zPublisherArticle
http://rdcu.be/t7pRPublisherFree ReadCube access
Additional Information:© 2013 Springer Science+Business Media New York. Published online: 29 June 2013. This work was supported by a subcontract from Colorado State University under NIH contract HHSN 266200400091c. The authors thank Dr. Larry Dangott of the Protein Chemistry Lab at Texas A&M University for mass spectrometry analysis.
Funders:
Funding AgencyGrant Number
Colorado State UniversityUNSPECIFIED
NIHHHSN 266200400091c
Subject Keywords:Th-17; IL-17A; Guinea pig; Protein
Issue or Number:3
Record Number:CaltechAUTHORS:20140110-140009344
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20140110-140009344
Official Citation:Dirisala, V., Jeevan, A., Ramasamy, S., & McMurray, D. (2013). Molecular Cloning, Expression, and In Silico Structural Analysis of Guinea Pig IL-17. Molecular Biotechnology, 55(3), 277-287. doi: 10.1007/s12033-013-9679-z
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:43316
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:21 Jan 2014 20:32
Last Modified:03 Oct 2019 06:06

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