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Identification and Molecular Characterization of Microneme 5 of Eimeria acervulina
In the present study, the microneme 5 gene of Eimeria acervulina (E. acervulina) (EaMIC5) was cloned and characterized. Specific primers for the rapid amplification of cDNA ends (RACE) were designed based on the expressed sequence tag (EST, GenBank Accession No. EH386430.1) to amplify the 3′- and 5′...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4274027/ https://www.ncbi.nlm.nih.gov/pubmed/25531898 http://dx.doi.org/10.1371/journal.pone.0115411 |
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author | Zhang, ZhenChao Huang, JingWei Li, MengHui Sui, YuXia Wang, Shuai Liu, LianRui Xu, LiXin Yan, RuoFeng Song, XiaoKai Li, XiangRui |
author_facet | Zhang, ZhenChao Huang, JingWei Li, MengHui Sui, YuXia Wang, Shuai Liu, LianRui Xu, LiXin Yan, RuoFeng Song, XiaoKai Li, XiangRui |
author_sort | Zhang, ZhenChao |
collection | PubMed |
description | In the present study, the microneme 5 gene of Eimeria acervulina (E. acervulina) (EaMIC5) was cloned and characterized. Specific primers for the rapid amplification of cDNA ends (RACE) were designed based on the expressed sequence tag (EST, GenBank Accession No. EH386430.1) to amplify the 3′- and 5′-ends of EaMIC5. The full length cDNA of this gene was obtained by overlapping the sequences of 3′- and 5′-extremities and amplification by reverse transcription PCR. Sequence analysis revealed that the open reading frame (ORF) of EaMIC5 was 336 bp and encoded a protein of 111 amino acids with 12.18 kDa. The ORF was inserted into pET-32a (+) to produce recombinant EaMIC5. Using western blotting assay, the recombinant protein was successfully recognized by the sera of chicks experimentally infected with E. acervulina, while the native protein in the somatic extract of sporozoites was as well detected by sera from rats immunized with the recombinant protein of EaMIC5. Immunofluorescence analysis using antibody against recombinant protein EaMIC5 indicated that this protein was expressed in the sporozoites and merozoites stages of E. acervulina. Animal challenge experiments demonstrated that the recombinant protein of EaMIC5 could significantly increase the average body weight gains, decrease the mean lesion scores and the oocyst outputs of the immunized chickens, and presented anti-coccidial index (ACI) more than 160. All the above results suggested that the EaMIC5 was a novel E. acervulina antigen and could be an effective candidate for the development of a new vaccine against this parasite. |
format | Online Article Text |
id | pubmed-4274027 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-42740272014-12-31 Identification and Molecular Characterization of Microneme 5 of Eimeria acervulina Zhang, ZhenChao Huang, JingWei Li, MengHui Sui, YuXia Wang, Shuai Liu, LianRui Xu, LiXin Yan, RuoFeng Song, XiaoKai Li, XiangRui PLoS One Research Article In the present study, the microneme 5 gene of Eimeria acervulina (E. acervulina) (EaMIC5) was cloned and characterized. Specific primers for the rapid amplification of cDNA ends (RACE) were designed based on the expressed sequence tag (EST, GenBank Accession No. EH386430.1) to amplify the 3′- and 5′-ends of EaMIC5. The full length cDNA of this gene was obtained by overlapping the sequences of 3′- and 5′-extremities and amplification by reverse transcription PCR. Sequence analysis revealed that the open reading frame (ORF) of EaMIC5 was 336 bp and encoded a protein of 111 amino acids with 12.18 kDa. The ORF was inserted into pET-32a (+) to produce recombinant EaMIC5. Using western blotting assay, the recombinant protein was successfully recognized by the sera of chicks experimentally infected with E. acervulina, while the native protein in the somatic extract of sporozoites was as well detected by sera from rats immunized with the recombinant protein of EaMIC5. Immunofluorescence analysis using antibody against recombinant protein EaMIC5 indicated that this protein was expressed in the sporozoites and merozoites stages of E. acervulina. Animal challenge experiments demonstrated that the recombinant protein of EaMIC5 could significantly increase the average body weight gains, decrease the mean lesion scores and the oocyst outputs of the immunized chickens, and presented anti-coccidial index (ACI) more than 160. All the above results suggested that the EaMIC5 was a novel E. acervulina antigen and could be an effective candidate for the development of a new vaccine against this parasite. Public Library of Science 2014-12-22 /pmc/articles/PMC4274027/ /pubmed/25531898 http://dx.doi.org/10.1371/journal.pone.0115411 Text en © 2014 Zhang et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Zhang, ZhenChao Huang, JingWei Li, MengHui Sui, YuXia Wang, Shuai Liu, LianRui Xu, LiXin Yan, RuoFeng Song, XiaoKai Li, XiangRui Identification and Molecular Characterization of Microneme 5 of Eimeria acervulina |
title | Identification and Molecular Characterization of Microneme 5 of Eimeria acervulina
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title_full | Identification and Molecular Characterization of Microneme 5 of Eimeria acervulina
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title_fullStr | Identification and Molecular Characterization of Microneme 5 of Eimeria acervulina
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title_full_unstemmed | Identification and Molecular Characterization of Microneme 5 of Eimeria acervulina
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title_short | Identification and Molecular Characterization of Microneme 5 of Eimeria acervulina
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title_sort | identification and molecular characterization of microneme 5 of eimeria acervulina |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4274027/ https://www.ncbi.nlm.nih.gov/pubmed/25531898 http://dx.doi.org/10.1371/journal.pone.0115411 |
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