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- W55882512 abstract "Tea caffeine synthase(TCS)gene cDNA code region was amplified by RT-PCR.The DNA fragment was cloned into expression vector pET 32a(+),forming recombinant pET-TCS.Expression of the plasmid in Escherichia coli BL21trxB(DE3)resulted in the production of a fusion protein(62 000)consisting of 112 amino acids of thioredoxin and 370 amino acids of TCS.With the inducing time increasing,the amount of fusion protein also increased,the highest content amounted to 39.14% of gross protein.Different temperature(15,25 and 37 ℃)could induce expression of fusion protein,and the higher temperature was,the more gross fusion protein and soluble fusion protein was produced.Most fusion protein was expressed in the cytoplasm and was soluble,a little of fusion protein was expressed in periplasm or expressed as inclusion bodies,and no fusion protein was found in culture.Furthermore,the results of in vitro enzymatic reaction suggested that the fusion protein had biological-activity and could catalyze the methylation of theobromine to caffeine." @default.
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- W55882512 date "2004-01-01" @default.
- W55882512 modified "2023-09-23" @default.
- W55882512 title "Expression of tea caffeine synthase cDNA in Escherichia coli" @default.
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