title: Evidence of alternative splicing of the chi2 chitinase gene from Metarhizium anisopliae.
abstract: Metarhizium anisopliae is a filamentous fungus used in the biological control of arthropods and produces several chitinases in order to break the host cuticle chitin fibers. Chitinase function during fungal cell development and/or infection processes is also an important aspect when analyzing the life cycle of entomopathogens. The expression profile analysis of the endochitinase chi2 gene acquired by RT-PCR experiments indicated the presence of two different transcripts, suggesting the occurrence of alternative splicing in the chi2 gene. The presence of two transcripts, characterized by the removal or retention of the second 72 bp intron, was further confirmed by DNA sequencing, Northern blot and qRT-PCR. Furthermore, we detected the synthesis of two different proteins from the transcripts by two-dimensional Western blot and mass spectrometry analyses. This is the first reported occurrence of alternative splicing in M. anisopliae.
authors.affiliation: Centro de Biotecnologia da Universidade Federal do Rio Grande do Sul, Programa de Pós-graduação em Biologia Celular e Molecular, Prédio 43-421, Bairro Agronomia, CEP 91501-970, Porto Alegre, Rio Grande do Sul, Brazil.
authors.forename: Juliano Tomazzoni
authors.initials: JT
authors.lastname: Boldo
authors.forename: Karina Bohrer
authors.initials: KB
authors.lastname: do Amaral
authors.forename: Angela
authors.initials: A
authors.lastname: Junges
authors.forename: Paulo Marcos
authors.initials: PM
authors.lastname: Pinto
authors.forename: Charley Christian
authors.initials: CC
authors.lastname: Staats
authors.forename: Marilene Henning
authors.initials: MH
authors.lastname: Vainstein
authors.forename: Augusto
authors.initials: A
authors.lastname: Schrank
mesh_terms: D017398:Alternative Splicing; D001483:Base Sequence; D015153:Blotting, Western; D002686:Chitin / Q000235:genetics / Q000378:metabolism; D002688:Chitinases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D005796:Genes; D052981:Metarhizium / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D002686:Chitin; D002688:Chitinases