Cloning, characterization and expression of glucoamylase gene from ectomycorrhizal basidomycete, Tricholoma matsutake
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- 2016.04.02
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- 2011.06
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서지정보
ㆍ발행기관 : 한국버섯학회
ㆍ수록지정보 : 한국버섯학회지 / 9권 / 2호
ㆍ저자명 : Jianing Wan, Ruirong Yi, Yan Li, Yukiko Kinjo, Aki Sadashima, Takao Terashita, Katsuji Yamanaka, Tadanori Aimi
목차
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
영어 초록
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.
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