Overexpression Vector Construction and Bioinformatics Analysis of Dt4CL from Dracocephalum tanguticum Maxim
1 Tibetan Collaborative Innovation Centre of Agricultural and Animal Husbandry Resources, Food Science College, Tibet Agriculture and Animal Husbandry University, Nyingchi, 860000
2 TAAHC-SWU Medicinal Plant Joint R&D Centre, Tibet Agricultural and Animal Husbandry University, Tibet 860000
3 Key Laboratory of Forest Ecology in Tibet Plateau (Tibet Agricultural & Animal Husbandry University), Ministry of Education, Tibet, 860000
Medicinal Plant Research, 2020, Vol. 10, No. 6
Received: 06 Sep., 2020 Accepted: 08 Sep., 2020 Published: 08 Sep., 2020
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This article was first published in Molecular Plant Breeding ( ISSN1672-416X，CN46-1068/S) in Chinese, and here was authorized to translate and publish the paper in English under the terms of Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
4-coumarate (4-coumarate: coenzyme A ligase, 4CL) is one of the key enzymes in the biosynthesis of flavonoids and lignin. It is located at the key point of the transition from the main pathway to the branch pathway of phenylalanine metabolism, and controls the entry into different secondary metabolic pathways to produce different secondary metabolites. In this experiment, a gene named Dt4CL1 was cloned from Dracocephalum tanguticum Maxim by PCR. The overexpression vector Dt4CL1-PHB was constructed by double enzyme digestion and plasmid recombination. The protein encoded by the gene was analyzed by bioinformatics, including physical and chemical properties, hydrophobicity, signal peptide, secondary structure, tertiary structure and phylogenetic tree. The results showed that ORF was 1 719 bp in length and encoded 572 amino acids. The predicted molecular weight was 61.61 kD and the theoretical PI was 5.39. Phylogenetic analysis showed that Dt4CL1 protein had the highest homology with the 4CL protein of Salvia miltiorrhiza and Prunella vulgaris. It provides a scientific basis for the study of the function of Dt4CL1.
Dracocephalum tanguticum Maxim; 4CL; Overexpression vector; Bioinformatics Analysis
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