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Construction of Magnaporthe grisea normalized cDNA library using SMART and recombinational cloning

Construction of Magnaporthe grisea normalized cDNA library using SMART and recombinational cloning

Phytopathology 93(6 Suppl): S40

ISSN/ISBN: 0031-949X

The rice blast fungus, Magnaporthe grisea, is one of the most studied plant pathogenic fungi. Whole genome sequence of this pathogen along with automated annotation and ORF prediction was released in June 2002. About 12,000 genes are predicted in the genome of M. grisea. EST alignment has been an essential tool to confirm gene expression, and contributes to the annotation. In addition, cDNA clones are useful for functional genomics, particularly proteome analysis. However, the EST projects often suffer from redundancy within cDNA libraries, which imposes a restraint in gene finding efforts. Various cDNA normalization strategies have been reported. We generated full-length cDNA library using SMART (Switching Mechanism At 5' end of the RNA Transcript) technology. This technique relies on the intrinsic TdT activity of reverse transcriptase to create full-length cDNA. The SMART technology can be integrated into recombinational cloning system such as the GATEWAY technology. We will discuss the use of SMART and GATEWAY technologies to construct a normalized full-length cDNA library from the appressorial stage of the rice blast fungus.

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Accession: 034641348

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