Cloning and Nucleotide Sequence of the Ribonuclease T_1 Gene (rntA) from Aspergillus oryzae and Its Expression in Saccharomyces cerevisiae and Aspergillus oryzae

概要

論文の詳細を見る
A genomic DNA encoding ribonuclease (RNase) T_1 from Aspergillus oryzae was cloned using a synthetic oligonucleotide probe. The cloned gene (designated rntA) encoded functional RNase T_1, since an A. oryzae transformant with multiple copies of the rntA gene showed higher RNase T_l activity (over 200 times) than a transformant with a vector. A cDNA was cloned by reverse transcription polymerase chain reaction (RT-PCR) with primers corresponding to the 5' terminus and 3' terminus of the reading frame of the rntA gene. Nucleotide sequencing analysis of both DNAs found that RNase T_1 had a prepro-sequence consisting of 26 amino acids and the rntA gene had only one intron (114 bp) in the region encoding the signal sequence. The A. oryzae transformant with cDNA controlled by the amyB promoter also showed higher activity (over 300 times), indicating that the cloned cDNA encoded functional RNase T_1. On the other hand, the Saccharomyces cerevisiae transformant with cDNA controlled by the GAL1 promoter could not grow on a medium containing galactose. These results suggests that A. oryzae may have a protection mechanism from RNase T_1.
社団法人日本農芸化学会の論文
1995-10-23