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Journal of Tea Science ›› 2006, Vol. 26 ›› Issue (4): 243-248.doi: 10.13305/j.cnki.jts.2006.04.002

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Construction of Tea Caffeine Synthase Gene RNAi Vector

ZHANG Guang-hui1, 2, LIANG Yue-rong1, *, LU Jian-liang1, DONG Jun-jie1   

  1. 1. Department of Tea Science, Zhejiang University, Hangzhou 310029, China;
    2. Xuzhou Higher Vocational School of Bioengineering, Xuzhou 221006, China
  • Received:2006-04-11 Revised:2006-09-13 Online:2006-12-25 Published:2019-09-11

Abstract: Tea Caffeine synthase (TCS) is one of the key enzymes involved in caffieine biosynthsis in tea plant (Camellia sinensis),which catalyses conversions of 7-methylxanthine to theobromine and theobromine to caffeine. Inhibition of TCS gene expression can leads to breeding low caffeine tea cultivars. Two cDNA fragments of TCS gene were amplified by RT-PCR, and ligated into T-vector. The two TCS gene fragments were inserted into RNAi vector pFGC5941 in reverse direction after double digestion with two pairs of restriction endonucleases. The insertion of two fragments, namely pFGC5941-TCS02 and pFGC5941-TCS03, into the RNAi vector were confirmed by PCR and DNA sequencing.

Key words: tea plant (Camellia sinensis), caffeine synthase, RNA interference, PCR

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