Expression of Citrus tristeza virus coat protein gene in Escherichia coli
Citrus tristeza virus (CTV) is among the most destructive pathogens of citrus and causes substantial economic losses in citrus-growing industry worldwide. Considering recent distribution of this pathogen and its capability of transmission by existing aphid vectors in Iran, detection of this virus is...
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| Format: | Article |
| Language: | English |
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University of Tarbiat Modares
2013-12-01
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| Series: | Journal of Crop Protection |
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| Online Access: | http://jcp.modares.ac.ir/article-3-12320-en.html |
| _version_ | 1818553486426505216 |
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| author | Amir Amiri Sadeghan Masoud Shams-Bakhsh Bagher Yakhchali |
| author_facet | Amir Amiri Sadeghan Masoud Shams-Bakhsh Bagher Yakhchali |
| author_sort | Amir Amiri Sadeghan |
| collection | DOAJ |
| description | Citrus tristeza virus (CTV) is among the most destructive pathogens of citrus and causes substantial economic losses in citrus-growing industry worldwide. Considering recent distribution of this pathogen and its capability of transmission by existing aphid vectors in Iran, detection of this virus is enforceable for controlling the damage caused by this pathogen in Iran, as one of the major citrus producing countries. Toward this aim, developing a reliable and sensitive detection method such as enzyme- linked immunosorbant assay (ELISA) would be the first step to detect CTV in large scale screenings of field samples. As the serological method requires great amounts of specific antibody, the consequent preparation of a large scale antigen source for immunization process is necessary. In this study the coat protein gene of CTV (CP25) was amplified by polymerase chain reaction from a cloned CP25 gene in pTZ57R/T and subcloned in pET26b expression vector and named pET-CP25. Two Escherichia coli strains of BL21 and Rosetta Gami (DE3) were transformed by pET-CP25. Expression of recombinant protein was induced by IPTG. The authenticity of recombinant protein was confirmed by western immunoblot analysis using a polyclonal antiserum against CTV particles. The results indicated that CTV coat protein gene was expressed in E.coli. This recombinant protein could be used as a source of antigen for immunization process. |
| first_indexed | 2024-12-12T09:26:26Z |
| format | Article |
| id | doaj.art-3fb637ebd2db4e6bbe64daea7e179bf4 |
| institution | Directory Open Access Journal |
| issn | 2251-9041 2251-905X |
| language | English |
| last_indexed | 2024-12-12T09:26:26Z |
| publishDate | 2013-12-01 |
| publisher | University of Tarbiat Modares |
| record_format | Article |
| series | Journal of Crop Protection |
| spelling | doaj.art-3fb637ebd2db4e6bbe64daea7e179bf42022-12-22T00:29:00ZengUniversity of Tarbiat ModaresJournal of Crop Protection2251-90412251-905X2013-12-0124387393Expression of Citrus tristeza virus coat protein gene in Escherichia coliAmir Amiri Sadeghan0Masoud Shams-Bakhsh1Bagher Yakhchali2 Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran. Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran. National Research Institute of Genetic Engineering and Biotechnology, Tehran. Iran. Citrus tristeza virus (CTV) is among the most destructive pathogens of citrus and causes substantial economic losses in citrus-growing industry worldwide. Considering recent distribution of this pathogen and its capability of transmission by existing aphid vectors in Iran, detection of this virus is enforceable for controlling the damage caused by this pathogen in Iran, as one of the major citrus producing countries. Toward this aim, developing a reliable and sensitive detection method such as enzyme- linked immunosorbant assay (ELISA) would be the first step to detect CTV in large scale screenings of field samples. As the serological method requires great amounts of specific antibody, the consequent preparation of a large scale antigen source for immunization process is necessary. In this study the coat protein gene of CTV (CP25) was amplified by polymerase chain reaction from a cloned CP25 gene in pTZ57R/T and subcloned in pET26b expression vector and named pET-CP25. Two Escherichia coli strains of BL21 and Rosetta Gami (DE3) were transformed by pET-CP25. Expression of recombinant protein was induced by IPTG. The authenticity of recombinant protein was confirmed by western immunoblot analysis using a polyclonal antiserum against CTV particles. The results indicated that CTV coat protein gene was expressed in E.coli. This recombinant protein could be used as a source of antigen for immunization process.http://jcp.modares.ac.ir/article-3-12320-en.htmlrecombinant proteinwestern blot analysiselisa |
| spellingShingle | Amir Amiri Sadeghan Masoud Shams-Bakhsh Bagher Yakhchali Expression of Citrus tristeza virus coat protein gene in Escherichia coli Journal of Crop Protection recombinant protein western blot analysis elisa |
| title | Expression of Citrus tristeza virus coat protein gene in Escherichia coli |
| title_full | Expression of Citrus tristeza virus coat protein gene in Escherichia coli |
| title_fullStr | Expression of Citrus tristeza virus coat protein gene in Escherichia coli |
| title_full_unstemmed | Expression of Citrus tristeza virus coat protein gene in Escherichia coli |
| title_short | Expression of Citrus tristeza virus coat protein gene in Escherichia coli |
| title_sort | expression of citrus tristeza virus coat protein gene in escherichia coli |
| topic | recombinant protein western blot analysis elisa |
| url | http://jcp.modares.ac.ir/article-3-12320-en.html |
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