In order to confirm the presence of putative glucoamylase gene in Tricholoma matsutake genome, the genomic DNA was prepared from T. matsutake NBRC30773 strain and was used as template to clone the glucoamylases gene (TmGlu1). We obtained the nucleotide sequence of TmGlu1 and its franking region. The coding region (from ATG to stop codon) is 2,186 bp. The locations of exons and introns were determined from the nucleotide sequences of 3’- and 5’-RACE PCR and RT- PCR products. On the other hand, to investigate the relationship between composition of medium and glucoamylase expression, we checked the expression level of glucoamylase gene by realtime reverse transcription PCR and measurement of glucoamylase enzyme activity. It was found that enzyme activity of glucoamylase was very low in different medium. Expression of glucoamylases gene appeared to not be affected by different carbon source.

ABSTRACT
 Introduction
 Materials and method
  Medium composition and culture conditions
  DNA and RNA preparation
  Amplification of glucoamylase genes
  DNA sequencing and computer analysis of nucleotideand protein sequences
  Reverse transcription-PCR (RT-PCR) procedure
  Glucoamylase assays
  Total protein assays
  Real-time PCR assay
 Results
 Discussion
 Acknowledgments
 References

• Jianing Wan(Faculty of Agriculture, Tottori University, 4-101 Koyamacho Minami, Tottori 680-8553, Japan)
• Ruirong Yi( Faculty of Agriculture, Tottori University, 4-101 Koyamacho Minami, Tottori 680-8553, Japan) | Ruirong Yi
• Yan Li( Faculty of Agriculture, Tottori University, 4-101 Koyamacho Minami, Tottori 680-8553, Japan) | Yan Li
• Yukiko Kinjo( Faculty of Agriculture, Tottori University, 4-101 Koyamacho Minami, Tottori 680-8553, Japan) | Yukiko Kinjo
• Aki Sadashima( Faculty of Agriculture, Tottori University, 4-101 Koyamacho Minami, Tottori 680-8553, Japan) | Aki Sadashima
• Takao Terashita( Faculty of Agriculture, Kinki University, 3327-204, Nakamachi, Nara 631-8505, Japan) | Takao Terashita
• Katsuji Yamanaka( Kyoto Mycological Institute, 1-55 Misasahirabayashi, Yamashina, Kyoto 607-8406, Japan) | Katsuji Yamanaka
• Tadanori Aimi( Faculty of Agriculture, Tottori University, 4-101 Koyamacho Minami, Tottori 680-8553, Japan) | Tadanori Aimi