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Bulletin of Botanical Research ›› 2013, Vol. 33 ›› Issue (1): 58-65.doi: 10.7525/j.issn.1673-5102.2013.01.010

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Cloning of Camellia nitidissima Flavonol Synthase cDNA and Construction of Sense,RNA Interference Expression Vectors

ZHOU Xing-Wen;LI Ji-Yuan*;Yin Heng-Fu;FAN Zheng-Qi;Li Xin-Lei   

  1. 1.Research Institute of Subtropical Forestry,Chinese Academy of Forestry,Fuyang 311400;2.Yulin Normal University,Yulin 537000
  • Received:1900-01-01 Revised:1900-01-01 Online:2013-01-20 Published:2013-01-20
  • Contact: LI Ji-Yuan
  • Supported by:

Abstract: A full-length cDNA sequence of flavonol synthase (FLS) gene was obtained from petals of Camellia nitidissima using the methods of Reverse Transcription PCR and RACE, and named CnFLS(GenBank accession No.JF343560.1). The sense expression vector pCAM-CnFLS was constructed successfully while the gene FLS amplified by the primers P3/P4 and inserted into PMD18-T vector was connected to expression vector pCAMBIA1300. The RNA interference (RNAi) expression vector was established by many times enzyme digestion and connection on the base of intermediate vector pUCCRNAi, expression vector pCAMBIA1300 and the combined vector PMD18-T with a conserved segment of 250 bp which was amplified by primers G1 and G2. The successful construction of sense and RNAi expression vectors provides a foundation for further studying on CnFLS gene functions and its effects to flower color.

Key words: Camellia nitidissima Chi, flavonol synthase, RNA interference, construction of expression vector

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