Evidence of alternative splicing of the chi2 chitinase gene from Metarhizium anisopliae.
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.
Boldo
Juliano Tomazzoni
JT
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.
do Amaral
Karina Bohrer
KB
Junges
Angela
A
Pinto
Paulo Marcos
PM
Staats
Charley Christian
CC
Vainstein
Marilene Henning
MH
Schrank
Augusto
A
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
D002686:Chitin; D002688:Chitinases