Pilot scale genome sequencing of Aspergillus nidulans and cDNA sequencing of Aspergillus oryzae.

Masavuki Machida, Mari Nakagawa, Sumiko Kunihiro, Kumiko Takase, Makoto Yasukaw, and Mariko Manabe.


In the course of world wide effort to complete the genome sequence of Aspergillus nidulans, we started the pilot scale sequencing of a part of the ordered cosmids library. We picked the cosmids locating in the middle part of chromosome VIII and neighboring to the cosmid which has been sequenced by Prade et al. The sequencing was done mainly by the primer walking method using the internal-labeling protocol and analyzed by Li-Cor model 4200L DNA sequencer. Since longer than 800 nucleotide sequence could be analyzed in a single run, approximately 160 reactions were expected to complete the sequence of both strands of cosmid's inserts. We examined the condition to adapt the internal-labeling protocol to cosmid sequencing and found that the successful long-read sequencing depended on higher concentration of IRD-labeled dATP, optimization of cosmid amount and the higher temperature for denaturation step. We have initiated the random cDNA sequencing of Aspergillus oryzae cDNA libraries prepared from the cells grown in a rich medium and in the starved condition. We are preparing the database of the above sequence data on our Web server, which will be soon available.

abstract No: 

Fungal Genet. Newsl. 46 (Supl):

Full conference title: 

Fungal Genetics Conference 20th
    • Fungal Genetics Conference 20th (1999)