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Cloning, Expression, and Homology Modeling of GroEL Protein from Leptospira interrogans Serovar Autumnalis Strain N2
Leptospirosis is an infectious bacterial disease caused by Leptospira species. In this study, we cloned and sequenced the gene encoding the immunodominant protein GroEL from L. interrogans serovar Autumnalis strain N2, which was isolated from the urine of a patient during an outbreak of leptospirosi...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Elsevier
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5054446/ https://www.ncbi.nlm.nih.gov/pubmed/22196358 http://dx.doi.org/10.1016/S1672-0229(11)60018-1 |
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author | Natarajaseenivasan, Kalimuthusamy Shanmughapriya, Santhanam Velineni, Sridhar Artiushin, Sergey C. Timoney, John F. |
author_facet | Natarajaseenivasan, Kalimuthusamy Shanmughapriya, Santhanam Velineni, Sridhar Artiushin, Sergey C. Timoney, John F. |
author_sort | Natarajaseenivasan, Kalimuthusamy |
collection | PubMed |
description | Leptospirosis is an infectious bacterial disease caused by Leptospira species. In this study, we cloned and sequenced the gene encoding the immunodominant protein GroEL from L. interrogans serovar Autumnalis strain N2, which was isolated from the urine of a patient during an outbreak of leptospirosis in Chennai, India. This groEL gene encodes a protein of 60 kDa with a high degree of homology (99% similarity) to those of other leptospiral serovars. Recombinant GroEL was overexpressed in Escherichia coli. Immunoblot analysis indicated that the sera from confirmed leptospirosis patients showed strong reactivity with the recombinant GroEL while no reactivity was observed with the sera from seronegative control patient. In addition, the 3D structure of GroEL was constructed using chaperonin complex cpn60 from Thermus thermophilus as template and validated. The results indicated a Z-score of −8.35, which is in good agreement with the expected value for a protein. The superposition of the Cα traces of cpn60 structure and predicted structure of leptospiral GroEL indicates good agreement of secondary structure elements with an RMSD value of 1.5 Å. Further study is necessary to evaluate GroEL for serological diagnosis of leptospirosis and for its potential as a vaccine component. |
format | Online Article Text |
id | pubmed-5054446 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-50544462016-10-14 Cloning, Expression, and Homology Modeling of GroEL Protein from Leptospira interrogans Serovar Autumnalis Strain N2 Natarajaseenivasan, Kalimuthusamy Shanmughapriya, Santhanam Velineni, Sridhar Artiushin, Sergey C. Timoney, John F. Genomics Proteomics Bioinformatics Article Leptospirosis is an infectious bacterial disease caused by Leptospira species. In this study, we cloned and sequenced the gene encoding the immunodominant protein GroEL from L. interrogans serovar Autumnalis strain N2, which was isolated from the urine of a patient during an outbreak of leptospirosis in Chennai, India. This groEL gene encodes a protein of 60 kDa with a high degree of homology (99% similarity) to those of other leptospiral serovars. Recombinant GroEL was overexpressed in Escherichia coli. Immunoblot analysis indicated that the sera from confirmed leptospirosis patients showed strong reactivity with the recombinant GroEL while no reactivity was observed with the sera from seronegative control patient. In addition, the 3D structure of GroEL was constructed using chaperonin complex cpn60 from Thermus thermophilus as template and validated. The results indicated a Z-score of −8.35, which is in good agreement with the expected value for a protein. The superposition of the Cα traces of cpn60 structure and predicted structure of leptospiral GroEL indicates good agreement of secondary structure elements with an RMSD value of 1.5 Å. Further study is necessary to evaluate GroEL for serological diagnosis of leptospirosis and for its potential as a vaccine component. Elsevier 2011-10 2011-12-23 /pmc/articles/PMC5054446/ /pubmed/22196358 http://dx.doi.org/10.1016/S1672-0229(11)60018-1 Text en © 2011 Beijing Institute of Genomics http://creativecommons.org/licenses/by-nc-sa/3.0/ This is an open access article under the CC BY-NC-SA license (http://creativecommons.org/licenses/by-nc-sa/3.0/). |
spellingShingle | Article Natarajaseenivasan, Kalimuthusamy Shanmughapriya, Santhanam Velineni, Sridhar Artiushin, Sergey C. Timoney, John F. Cloning, Expression, and Homology Modeling of GroEL Protein from Leptospira interrogans Serovar Autumnalis Strain N2 |
title | Cloning, Expression, and Homology Modeling of GroEL Protein from Leptospira interrogans Serovar Autumnalis Strain N2 |
title_full | Cloning, Expression, and Homology Modeling of GroEL Protein from Leptospira interrogans Serovar Autumnalis Strain N2 |
title_fullStr | Cloning, Expression, and Homology Modeling of GroEL Protein from Leptospira interrogans Serovar Autumnalis Strain N2 |
title_full_unstemmed | Cloning, Expression, and Homology Modeling of GroEL Protein from Leptospira interrogans Serovar Autumnalis Strain N2 |
title_short | Cloning, Expression, and Homology Modeling of GroEL Protein from Leptospira interrogans Serovar Autumnalis Strain N2 |
title_sort | cloning, expression, and homology modeling of groel protein from leptospira interrogans serovar autumnalis strain n2 |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5054446/ https://www.ncbi.nlm.nih.gov/pubmed/22196358 http://dx.doi.org/10.1016/S1672-0229(11)60018-1 |
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