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Cloning of carnation GA 20-oxidase and the construction of a plant RNAi vector

Published online by Cambridge University Press:  30 October 2009

Jin Liang ,
Liu Yun ,
Sun Zhen-Yuan  and
Li Tian-Hong
Jin Liang
Affiliation:
Department of Fruit Science, College of Agronomy and Biotechnology, China Agricultural University/Key Laboratory of Stress Physiology and Molecular Biology for Tree Fruits of Beijing, Beijing 100193China
Liu Yun
Affiliation:
Department of Fruit Science, College of Agronomy and Biotechnology, China Agricultural University/Key Laboratory of Stress Physiology and Molecular Biology for Tree Fruits of Beijing, Beijing 100193China
Sun Zhen-Yuan
Affiliation:
The Research Institute of Flowers, Chinese Academy of Forestry, Beijing 100093, China
Li Tian-Hong*
Affiliation:
Department of Fruit Science, College of Agronomy and Biotechnology, China Agricultural University/Key Laboratory of Stress Physiology and Molecular Biology for Tree Fruits of Beijing, Beijing 100193China
*
*Corresponding author. E-mail: [email protected]
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Abstract

A pair of degenerate primers was designed based on a conserved domain of GA 20-oxidase reported in other plants. The full-length (1179 bp) carnation (Dianthus caryophyllus L. cv. Master) GA 20-oxidase cDNA (named Dc20ox) was cloned by reverse transcriptase-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). BLAST analysis revealed that the deduced amino acid sequence had high homology (66–75%) with the GA 20-oxidase sequences from other plants. An RNAi vector (pART400) was constructed from a 400 bp fragment representing a highly conserved region of GA 20-oxidase.

Keywords

carnationGA 20-oxidaseRNAi
Type
Research Papers
Information
Chinese Journal of Agricultural Biotechnology , Volume 6 , Issue 2 , August 2009 , pp. 127 - 134
Copyright
Copyright © China Agricultural University 2009

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