Molecular Cloning, Characterization and Expression Analysis of a Gene Encoding Hydroxyphenylpyruvate Reductase Involved
In this study, a novel full-length cDNA of hydroxyphenylpyruvate reductase (HPPR) involved in rosmarinic acid (RA) biosynthesis pathway in Perilla frutescensis is cloned (designated as PfHPPR) for the first time by conserved fragment PCR and rapid amplifi
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Molecular Cloning, Characterization and Expression Analysis of a Gene Encoding Hydroxyphenylpyruvate Reductase Involved in Rosmarinic Acid Biosynthesis Pathway from Perilla frutescens Xiaoling Lu, Lei Hao, Fang Wang and Chen Huang Abstract In this study, a novel full-length cDNA of hydroxyphenylpyruvate reductase (HPPR) involved in rosmarinic acid (RA) biosynthesis pathway in Perilla frutescensis is cloned (designated as PfHPPR) for the first time by conserved fragment PCR and rapid amplification of cDNA ends (RACE). The cDNA of PfHPPR is 1723 bp long with an open reading frame (ORF) of 939 bp encoding a protein of 313 amino acid residues. Bioinformatics analysis revealed that the deduced amino acid sequence of PfHPPR has high homology to the other known HPPR proteins. Quantitative real-time PCR analysis of the transcription pattern indicates that the PfHPPR expresses constitutively in all examined tissues but most highly in leaf. Besides, the expression of PfHPPR can be induced by several factors such as abscisic acid (ABA), salicylic acid (SA), and ultraviolet-B radiation (UV-B).
Keywords Perilla frutescens PfHPPR gene PCR Rosmarinic acid Sequence analysis
RACE Quantitative real-time
X. Lu (&) L. Hao F. Wang Laboratory of Food Additives and Nutrition, College of Food Engineering and Biological Technology, Tianjin University of Science and Technology, Tianjin 300457, People’s Republic of China e-mail: [email protected] C. Huang Tianjin Entry-Exit Inspection and Quarantine Bureau, Tianjin 300461, People’s Republic of China
T.-C. Zhang et al. (eds.), Proceedings of the 2012 International Conference on Applied Biotechnology (ICAB 2012), Lecture Notes in Electrical Engineering 251, DOI: 10.1007/978-3-642-37925-3_194, Ó Springer-Verlag Berlin Heidelberg 2014
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194.1 Introduction Perilla frutescens (PfHPPR) is an annual self-fertilizing plant belonging to the family Lamiaceae, which have been extensively recognized worldwide as a representative oil crop and traditional Chinese medicinal herb [1, 2]. The Perilla frutescens accumulates a large amount of essential oil on a dry weight basis, and most of which is composed of unsaturated fatty acid such as a-Linolenic acid and various kinds of secondary metabolites with high bioactivity [3–5]. One of these metabolites is RA, an ester of caffeic acid and 3, 4-dihydroxyphenyllactic acid which is most commonly found in Lamiaceae and Boraginaceae species, and also detected in other families of dicotyledonous and monocotyledonous plants as well as in some hornworts and ferns [6]. RA has a multitude of biological activities, such as antioxidant, antibacterial, anti-inflammatory, and antimutagen [7, 8], which makes it a very valuable product for pharmaceutical and cosmetic industries. The biosynthesis of RA (Fig. 194.1) had been reported since 1970 when Ellis and Towers found that the two aromatic amino acids L-phenylalanine and L-tyrosine were the precursors for this compound [9]. L-phenylalanine is transformed to 4-coumaroyl-CoA by t
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