Volume 16, Issue 3 (9-2021)                   MGj 2021, 16(3): 211-218 | Back to browse issues page

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karimi goyjelo R, Abdollahi Mandoulakani B. Isolation and characterization of cinnamate- 4-hydroxylase gene promoter in Ocimum basilicum L.. MGj. 2021; 16 (3) :211-218
URL: http://mg.genetics.ir/article-1-1674-en.html
Urmia University
Abstract:   (266 Views)
Basil (occimum basilicum L.) essential oil is a rich source of phenylpropaniod compounds such as phenyl cinnamate, methyl chavicol, methyl eugenol and eugenol. Cinnamate-4-hydroxylase (C4H) is one of the important key genes involved in phenylpropanoid biosynthesis. Nowadays considering the importance of the promoter in genetic manipulation, gene transfer and the production of secondary metabolites in plants, identification of the promoter sequence and its constituent elements is necessary. Hence, in the current investigation, thermal asymmetric interlaced PCR (TAIL-PCR) was used to identify the upstream sequences of the C4H gene in O. basilicum L. In this method two series of primers differing in length and annealing temperature were applied to amplify the target sequence: three specific primers with high annealing temperature and different sequences and six random primers with a short length and low annealing temperature. TAIL-PCR was performed in three stages which in the second and third stages, diluted PCR product of the first and second stages were used respectively as the template. After electrophoresis of the PCR products and identification of the interested band, the fragments were extracted and purified from the agarose gel, and sequenced in both directions. Subsequent bioinformatic analysis of the sequenced fragment using PLACE, PLANT CARE, Softberry and Neural Network Promoter Prediction softwares revealed a sequence with a length of 794 bp. The TATA Box of the promoter was recognized around nucleotide -35 and the transcription initiation site was 170 nucleotides upstream of the initiation codon. A group of various putative functional regulatory elements such as CAAT box, W box, Gbox and I box were also identified in the promoter. Therefore, the promoter detected (after validation), may be used in the gene transfer and manipulation programs in basil for enhancement of the valuable metabolites.
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Type of Study: Applicable | Subject: Subject 01
Received: 2020/11/9 | Accepted: 2021/07/3 | Published: 2021/09/21

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