Development of new tools for studying gene function in fungi based on the Gateway system.
Genomic information of many fungi has been released but large scale functional genomic studies are still limited by a lack of high-throughput methods. The low rates of homologous recombination and low rates of transformation are limiting steps in filamentous fungi, but the molecular tools are also lagging behind. In this paper we describe two new high-throughput functional genomic tools for filamentous fungi that are based on the Gateway technology. One system is the Gateway RNAi vector for fungi that allows gene silencing in a high-throughput manner. The other system is a high-throughput deletion construct system. These systems were tested using the PAC1 gene of Colletotrichum gloeosporioides. Using these types of approaches, large scale functional genomics experiments can be performed in filamentous fungi.
Shafran
Hadas
H
Department of Genomics, Agricultural Research Organization, The Volcani Center, Bet Dagan 50250, Israel.
Miyara
Itay
I
Eshed
Ravit
R
Prusky
Dov
D
Sherman
Amir
A
D020231:Colletotrichum / Q000235:genetics; D005658:Fungi / Q000235:genetics*; D017353:Gene Deletion; D005822:Genetic Vectors / Q000235:genetics; D016681:Genome, Fungal / Q000235:genetics; D023281:Genomics / Q000379:methods*; D008957:Models, Genetic; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction