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cDNA cloning and prokaryotic expression of β-glucosidase in tea plant [Camellia sinensis (L.) O. Kutze]

Published online by Cambridge University Press:  13 June 2008

Jiang Chang-Jun ,
Li Yuan-Hua  and
Fang Wan-Ping
Jiang Chang-Jun*
Affiliation:
Key Laboratory of Tea Biochemistry and Biotechnology, Ministry of Agriculture, Anhui Agricultural University, Hefei 230036, China
Li Yuan-Hua
Affiliation:
Key Laboratory of Tea Biochemistry and Biotechnology, Ministry of Agriculture, Anhui Agricultural University, Hefei 230036, China
Fang Wan-Ping
Affiliation:
Key Laboratory of Tea Biochemistry and Biotechnology, Ministry of Agriculture, Anhui Agricultural University, Hefei 230036, China
*
*Corresponding author. Email: [email protected]
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Abstract

The β-glucosidase gene has important effects on alcoholic aroma precursors and insect resistance of the tea plant [Camellia sinensis (L.) O. Kutze]. The complete cDNA sequence of β-glucosidase of the tea plant was cloned; its full length was 1475 bp, and shared 40–60% similarity with corresponding parts of the nucleotide sequence of β-glucosidase gene from other plants. Its secondary structure contains 14.33% α-helix, 25.43% β-pleated sheet and many functional amino acid domains. The β-glucosidase gene was cloned into the pET-32a expression system and expressed at high-efficiently in Escherichia coli BL21 (DE3); the molecular weight of expressed fusion protein was 63 kDa. The results of enzymic reaction showed that the fusion protein possessed normal bioactivity, and it could catalyse the dehydration of the glycosidic bond. The soluble fusion protein was expressed mainly in the cytoplasm.

Keywords

tea (Camellia sinensis)β-glucosidase; cDNAprokaryotic expression
Type
Research Article
Information
Chinese Journal of Agricultural Biotechnology , Volume 2 , Issue 2 , August 2005 , pp. 107 - 111
Copyright
Copyright © China Agricultural University and Cambridge University Press 2005

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