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Caracterização do gene LIP2 de Trichoderma harzianum

Authors: Vaz, Madalena;

Caracterização do gene LIP2 de Trichoderma harzianum

Abstract

Os fungos do género Trichoderma abrangem um grupo de fungos extremamente comuns em solos de todas as zonas climáticas. São eficientes produtores de enzimas com aplicações industriais, ou na natureza, estando envolvidos na degradação da parede celular dos fitopatogénios bem como na degradação de outros fungos, matéria orgânica e nutrientes secretados pelas raízes. As estirpes de Trichoderma produzem enzimas extracelulares e antibióticos com efeitos antifúngicos, sobretudo T. harzianum, T. virens e T. viride, sendo por isso usados como agentes de biocontrolo. De entre as enzimas de degradação das paredes celulares produzidas, encontram-se β-1,3 e β-1,6 glucanases, quitinases e proteases. Pensa-se que também as lipases poderão estar envolvidas na actividade enzimática de biocontrolo. Este trabalho teve como objectivo a caracterização do gene lip2 de Trichoderma harzianum, para isso recorreu-se a um conjunto de ferramentas de biologia molecular, bem como o recurso a programas bioinformáticos, de forma a efectuar a caracterização do gene, contribuindo assim para um melhor conhecimento deste. Procedeu-se à clonagem do gene lip2 num sistema de expressão, usando o vector pET-28a(+), e à avaliação da expressão da proteína lip2 por gel de SDS-PAGE em diferentes tempos de indução. Os resultados obtidos na clonagem indicaram uma banda total de 6584pb o que comprovou o sucesso da clonagem. A expressão da proteína após 8 horas de indução manifestou-se pela presença de uma banda de peso molecular estimado de 44kDa, observada por gel SDS-PAGE.T

richoderma spp. covers a group of fungi extremely common in soils of all climatic areas. These fungi are efficient enzyme producers, with indústrial applications, or in nature, involved in the degradation of the cell wall of the phytopathogens as well as in the degradation of other fungi, organic matter and nutrients secreted by roots. Trichoderma strains produce extracellular enzymes and antibiotics with antifungal effects, mainly T. harzianum, T. virens and T. viride, being by this way used as biocontrol agents. Among the degradation enzymes of the cells walls produced, are β-1,3 and β-1,6 glucanases, chitinases and proteases. Lipases could also be involved in enzymatic activity related with biocontrol. This work intention was the characterization of the lip2 gene of the T. harzianum, thus were used a set of tools from molecular biology and bioinformatics tools in order to perform the study, contributing for a better knowledge of the gene. Lip2 gene was cloned in an expression system, using the pET-28a(+) vector, and the expression evaluation of the lip2 protein was made in a SDS-PAGE gel at different induction times. The obtained results in the cloning showed a 6584bp band indicating that the cloning was well succeeded. The protein expression was verified at 8 hours of induction by the presence of a band with an estimated molecular weight of 44kDa, in a SDS-PAGE gel.

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Portugal
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Keywords

Trichoderma harzianum, Clonagem, Lipase, lip2, SDS-PAGE

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
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