A gene trap vector for Aspergillus oryzae was constructed for use in comprehensive analysis of gene function. This vector, pPTR-EGFP1, has the egfp gene as a reporter gene preceded by a splice acceptor sequence, followed by A. nidulans sC gene terminator and pyrithiamine resistant gene (ptrA) as a selective marker in A.oryzae. This vector was integrated into a host genome randomly to create tag-line transformants. Approximately 300 pyrithiamine-resistant transformants were thus obtained, one of which trapped the 5' end of the coding region of yeast DUR3 homologue. Our results showed that pPTR-EGFP1 could be a useful tool in the gene function analysis of A.oryzae, namely, by isolating unknown genes by plasmid rescue and by trapping the endogenous promoter activity.
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麹菌用ジーントラップベクター, pPTR-EGFP1の作成
nfri_report_No67p33-38p.pdf
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