Publication
Cloning and expression analysis of an endo‑1,3‑β‑d‑glucosidase from Phytophthora cinnamomi
| dc.contributor.author | Costa, Rodrigo Arthur Fonseca | |
| dc.contributor.author | Choupina, Altino | |
| dc.contributor.author | Dominguez, Ángel | |
| dc.date.accessioned | 2020-01-22T09:22:34Z | |
| dc.date.available | 2020-01-22T09:22:34Z | |
| dc.date.issued | 2019 | |
| dc.description.abstract | Phytophthora is considered one of the most destructive genus for many agricultural plant species worldwide, with a strong environmental and economic impact. Phytophthora cinnamomi is a highly aggressive Phytophthora species associated with the forest decline and responsible for the ink disease in chestnut trees (Castanea sativa Miller), a culture which is extremely important in Europe. This pathogenicity occurs due to the action of several enzymes like the hydrolysis of 1,3-β-glucans at specific sites by the enzyme endo-1,3-β-d-glucosidase. The aim of this work to analyze the heterologous expression in two microorganisms, Escherichia coli and Pichia pastoris, of an endo-1,3-β-d-glucosidase encoded by the gene ENDO1 (AM259651) from P. cinnamomi. Different plasmids were used to clone the gene on each organism and the real-time quantitative polymerase chain reaction was used to determine its level of expression. Homologous expression was also analyzed during growth in different carbon sources (glucose, cellulose, and sawdust) and time-course experiments were used for endo-1,3-β-d-glucosidase production. The highest expression of the endo-1,3-β-d-glucosidase gene occurred in glucose after 8 h of induction. In vivo infection of C. sativa by P. cinnamomi revealed an increase in endo-1,3-β-d-glucosidase expression after 12 h. At 24 h its expression decreased and at 48 h there was again a slight increase in expression, and more experiments in order to further explain this fact are underway. | pt_PT |
| dc.description.version | info:eu-repo/semantics/publishedVersion | pt_PT |
| dc.identifier.citation | Costa, Rodrigo; Choupina, Altino; Domínguez, Angel (2019). Cloning and expression analysis of an endo‑1,3‑β‑d‑glucosidase from Phytophthora cinnamomi. Molecular Biology Reports. ISSN 1573-4978. 47:2, p. 935-942 | pt_PT |
| dc.identifier.doi | 10.1007/s11033-019-05185-9 | pt_PT |
| dc.identifier.issn | 0301-4851 | |
| dc.identifier.uri | http://hdl.handle.net/10198/20462 | |
| dc.language.iso | eng | pt_PT |
| dc.peerreviewed | yes | pt_PT |
| dc.publisher | Springer | pt_PT |
| dc.rights.uri | http://creativecommons.org/licenses/by-nc/4.0/ | pt_PT |
| dc.subject | Castanea sativa | pt_PT |
| dc.subject | Endo-1,3-β-d-glucosidase | pt_PT |
| dc.subject | Heterologous expression | pt_PT |
| dc.subject | Homologous expression | pt_PT |
| dc.subject | Phytophthora cinnamomi | pt_PT |
| dc.subject | RT-qPCR | pt_PT |
| dc.title | Cloning and expression analysis of an endo‑1,3‑β‑d‑glucosidase from Phytophthora cinnamomi | pt_PT |
| dc.type | journal article | |
| dspace.entity.type | Publication | |
| oaire.citation.conferencePlace | Springer Nature Switzerland AG. Part of Springer Nature. | pt_PT |
| oaire.citation.endPage | 942 | pt_PT |
| oaire.citation.startPage | 935 | pt_PT |
| oaire.citation.title | Molecular Biology Reports | pt_PT |
| person.familyName | Choupina | |
| person.givenName | Altino | |
| person.identifier | 587972 | |
| person.identifier.ciencia-id | 1A14-77FC-9656 | |
| person.identifier.orcid | 0000-0002-3956-9398 | |
| person.identifier.rid | L-5382-2014 | |
| person.identifier.scopus-author-id | 14051602500 | |
| rcaap.rights | openAccess | pt_PT |
| rcaap.type | article | pt_PT |
| relation.isAuthorOfPublication | 314b895b-a5e2-4a92-9bc6-dd708c819be4 | |
| relation.isAuthorOfPublication.latestForDiscovery | 314b895b-a5e2-4a92-9bc6-dd708c819be4 |