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Previous issue date: 2009-02-27 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The genome of all organisms is subject to injuries that can be caused by endogenous and environmental factors. If these lesions are not corrected, it can be fixed generating a mutation which can be lethal to the organisms. In order to prevent this, there are different DNA repair mechanisms. These mechanisms are well known in bacteria, yeast, human, but not in plants. Two plant models Oriza sativa and Arabidopsis thaliana had the genome sequenced and due
to this some DNA repair genes have been characterized. The aim of this work is to characterized two sugarcane cDNAs that had homology to AP endonuclease: scARP1 and
scARP3. In silico has been done with these two sequences and other from plants. It has been observed domain conservation on these sequences, but the cystein at 65 position that is a characteristic from the redox domain in APE1 protein was not so conservated in plants. Phylogenetic relationship showed two branches, one branch with dicots and monocots sequence and the other branch with only monocots sequences. Another approach in order to
characterized these two cDNAs was to construct overexpression cassettes (sense and antisense orientation) using the 35S promoter. After that, these cassettes were transferred to the binary vector pPZP211. Furthermore, previously in the laboratory was obtained a plant from
nicotiana tabacum containing the overexpression cassette in anti-sense orientation. It has been observed that this plant had a slow development and problems in setting seeds. After
some manual crossing, some seeds were obtained (T2) and it was analyzed the T2 segregation. The third approach used in this work was to clone the promoter region from these
two cDNAs by PCR walking. The sequences obtained were analyzed using the program PLANTCARE. It was observed in these sequences some motives that may be related to
oxidative stress response / O genoma de todos os organismos est? sujeitos a les?es que podem ser causados por fatores end?genos e ambientais. Uma vez no genoma, as les?es podem levar a forma??o e ac?mulo de muta??es, as quais podem ser prejudiciais ao desenvolvimento do organismo. As vias de reparo de DNA s?o um mecanismo que permite o organismo detectar e corrigir essas les?es ou minimizar seus efeitos. V?rias vias de reparo de DNA s?o conhecidas e bem caracterizadas em animais e microorganismos. Em plantas, as vias de reparo ainda n?o s?o bem caracterizadas, mas muitas pesquisas t?m revelado a participa??o de todas as vias conhecidas no reparo do genoma das plantas, sendo os modelos mais estudados Arabidopsis thaliana e Oriza sativa. A via de reparo de interesse deste trabalho ? a via de BER, a qual apresenta v?rias prote?nas atuando no reparo do DNA. Por?m, a classe de prote?nas da via BER focadas s?o as AP endonucleases, respons?veis pela hidr?lise do s?tio AP. Este trabalho teve como objetivo caracterizar dois cDNAs de cana-de-a??car: scARP1 e scARP3, hom?logos a AP endonucleases de A.thaliana e identificados num trabalho de data-mining do projeto SUCEST. Para tanto, foram constru?dos cassetes de super-express?o, contendo o cDNA scARP1 sob o controle do promotor forte 35S. Al?m disso, anteriormente no laborat?rio foi obtido uma planta de icotiana tabacum contendo o cassete de super-express?o (35S+scARP1) em orienta??o anti-senso. Foi analisado o desenvolvimento desta planta, e foram obtidas tamb?m algumas sementes (T2) desta planta. Estas sementes foram germinadas e analisadas quanto ? presen?a do cassete de super-express?o e desenvolvimento. Al?m disso, para estes dois genes foram clonados as regi?es promotoras por PCR walking. Os fragmentos obtidos foram clonados, sequenciados e, analisados por meio do programa PLANTCARE. Os motivos encontrados nessas regi?es dos genes de cana-de-a??car foram comparados com potenciais regi?es promotoras das plantas de Arabidopis, O. sativa e S. bicolor. Estas an?lises mostraram a presen?a de diferentes motivos relacionados ? respostas ao estresse oxidativo
| Identifer | oai:union.ndltd.org:IBICT/oai:repositorio.ufrn.br:123456789/18626 |
| Date | 27 February 2009 |
| Creators | Oliveira, Andrea de Lima |
| Contributors | CPF:13451112825, http://lattes.cnpq.br/4808910380593455, Lima, Lucymara Fassarela Agnez, CPF:00297997750, http://lattes.cnpq.br/1083882171718362, Sluys, Marie-anne Van, CPF:80867359749, http://lattes.cnpq.br/5131787064674647, Scortecci, K?tia Castanho |
| Publisher | Universidade Federal do Rio Grande do Norte, Programa de P?s-Gradua??o em Gen?tica e Biologia Molecular, UFRN, BR, Gen?tica e Biologia Molecular |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | Portuguese |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Format | application/pdf |
| Source | reponame:Repositório Institucional da UFRN, instname:Universidade Federal do Rio Grande do Norte, instacron:UFRN |
| Rights | info:eu-repo/semantics/embargoedAccess |
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