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β-glucosidase cDNA Cloning in the Tea(Camellia sinensis ) and Its Prokaryotic Expression |
LI Yuan-Hua JIANG Chang-Jun** YANG Shun-Li YU You-Ben |
(Key Laboratory of Tea Biochemistry and Biotechnology, Ministry of Agriculture, Anhui Agricultural University, Hefei 230036, China) |
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Abstract Abstract:The β-glucosidase gene has important effects on the alcoholic aroma precursors and insect-resistance. The complete cDNA squence of β-glucosidase of tea(Camellia sinensis) was cloned and its full length was 1 475 bp (GenBank, Accession No. AF537127), and shared 40%~60% similarity to corresponding parts of β-glucosidase gene from other plants in nucleotide sequence. Its secondary structure contained 14.33% α-helical conformation, 25.43% β-sheet conformation and many function domains of amino acid. The β-glucosidase gene was cloned into the pET-32a expression vector and expressed high-efficiently in Escherichia coli BL21(DE3), and the molecular weight of expressed fusion protein was 63 kD. The results of emzymatic reaction showed that fusion protein possesed normal bioactivity, and it could catalyze the dehydration of the glycodic bond. The fusion protein was mainly expressed by soluble protein in cytoplasm.
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Received: 01 January 1900
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Corresponding Authors:
JIANG Chang-Jun
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