Molecular Cloning and Sequences Analysis of Chalcone Synthase Gene from Fagopyrum Tataricum
One of the most important key enzymes Chalcone Synthase (CHS) gene was cloned from Fagopyrum tataricum. Based on cDNA sequence conserved domain of CHS, a pair of primers were designed and used to amplify a fragment of FtCHS gene (GenBank accession: EU7152
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Molecular Cloning and Sequences Analysis of Chalcone Synthase Gene from Fagopyrum Tataricum Huangyuan She, Shaohong He, Zhi Zhou and Qitang Zhang Abstract One of the most important key enzymes Chalcone Synthase (CHS) gene was cloned from Fagopyrum tataricum. Based on cDNA sequence c onserved domain of CHS, a pair of primers were designed and used to amplify a fragment of FtCHS gene (GenBank accession: EU715255) using RT—PCR and RACE technique. A 1,463 bp full-length cDNA sequence was obtained. Analysis of CHS cDNA indicated that it encoded a peptide containing 393 amino acids. The sequence identity to palm leaf rhubarb (Rheumpahlmatum L.) species was all about 83 %. It is predicted that secondary structure of FtCHS contains α-helix (41.73 %), layers (16.28 %) and random coils (41.98 %); further research on three-dimensional modeling of FtCHS contains 11 α-helixes and 15 β-layers, which showed that protein was able to fold to a typical three-dimensional structure of CHS protein highly similar to that of MsCHS 2. The predicted tertiary structure demonstrated that FtCHS has combination sites. Keywords Fagopyrum tataricum • Chalcone synthase • Gene cloning • Sequence analysis
H. She (*) School of Chemistry, Biology and Agronomy, Anshun University, Anshun 561000, China e-mail: [email protected] S. He Ethnic teachers’ training school, Anshun 561000, China Z. Zhou Bureau of Education, Duyuan 558000, China H. She · Q. Zhang School of Life Sciences, Southwest University, Chongqing 400715, China
W. Du (ed.), Informatics and Management Science I, Lecture Notes in Electrical Engineering 204, DOI: 10.1007/978-1-4471-4802-9_85, © Springer-Verlag London 2013
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85.1 Introduction Fagopyrum tataricum (Linn.) Gaertn is an annual polygonaceae fagopyrum herbaceous plant which belongs to minor cereals and is mainly produced in highaltitude areas [1, 2]. Tartary buckwheat is rich in flavonoids compounds which have functions of lowering the blood sugar level and the blood fat and can be used to prevent and assist treating diseases such as diabetes mellitus, hyperlipidemia and high pressure [3]. Chalcone synthase (CHS, EC 2.3.1.47) is a polyketide synthase which catalyzes the first rate-limiting step of flavonoid biosynthesis, i.e., catalyzes termolecular malonyl-CoA and 4-coumaroyl-CoA and produces 4, 2′, 4′, 6′-tetrahydroxy chalcone [4, 5]. Genome sequence and cDNA sequence of CHS gene in some plants such as alfalfa (M. sativa) have been cloned and studied. The crystal structure of protein of alfalfa CHS 2 gene for prokaryotic expression has been reported [6], which has provided convenience for studying substrate specificity of CHS protein in vitro and hereditarily transform the biosynthesis of flavonoids compounds. In this research, in order to get the construction of buckwheat plant expression vector and transformation prepared, to realize the supersession of buckwheat flavonoid, and to establish the foundation for the achievement of new materials or new species with high flavonoid concentrati
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