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The Gene Cloning and Expression Analysis of C4H in Tea Plant (Camellia sinensis)

  • YAO Shengbo ,
  • WANG Wenzhao ,
  • LI Mingzhuo ,
  • XU Yujiao ,
  • WANG Yunsheng ,
  • LIU Yajun ,
  • GAO Liping ,
  • XIA Tao
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  • 1. Key Lab of Tea Biochemistry and Biotechnology, Ministry of Education, Anhui Agricultural University, Hefei 230036, China;
    2. School of Biology Science, Anhui Agricultural University, Hefei 230036, China

Received date: 2014-07-14

  Revised date: 2014-08-09

  Online published: 2019-08-23

Abstract

Cinnamate 4-hydroxylase (C4H) is a key enzyme in the phenylpropanoid pathway in tea plant. The gene can influence the biosynthesis of secondary metabolites such as lignin and flavonoids. The cDNA full-length of C4H gene was cloned from tea plant by rapid amplification of cDNA ends with a 1β518βbp open reading frame encoding a protein of 505 amino acids. The deduced protein molecular weight was 58.15βkD and its theoretical isoelectric point was 9.29. A 1β840βbp promoter sequence was isolated by genome walking method. The promoter region not only has the basic transcriptional elements of TATA-box and CAAT-box, but also has many potential inducible and tissue-specific cis-acting elements. Quantitative RT-PCR analysis showed that the CsC4H gene expressed in bud, leaf, stem and root. The gene was cloned into the expression vector pYES-DEST52 for eukaryotic expression in Saccharomyces cerevisiae WAT11. The enzyme reaction products were detected by LC-MS method. The results indicated that cinnamic acid was para-hydroxylated by target proteins to generate p-coumaric acid.

Cite this article

YAO Shengbo , WANG Wenzhao , LI Mingzhuo , XU Yujiao , WANG Yunsheng , LIU Yajun , GAO Liping , XIA Tao . The Gene Cloning and Expression Analysis of C4H in Tea Plant (Camellia sinensis)[J]. Journal of Tea Science, 2015 , 35(1) : 35 -44 . DOI: 10.13305/j.cnki.jts.2015.01.007

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