title: Characterization of Squalene synthase gene from Chlorophytum borivilianum (Sant. and Fernand.).
abstract: Saponins are important group of secondary metabolites known for their pharmacological properties. Chlorophytum borivilianum contains high amount of saponins and is thus, recognized as an important medicinal plant with aphrodisiac properties. Though the plant is well known for its pharmaceutical properties, there is meager information available about the genes and enzymes responsible for biosynthesis of saponins from this plant. Squalene synthase (SqS) is the key enzyme of saponin biosynthesis pathway and here, we report cloning and characterization of SqS gene from C. borivilianum. A full-length CbSqS cDNA consisting of 1,760 bp was cloned which contained an open reading frame (ORF) of 1,233 bp, encoding a protein of 411 amino acids. Analysis of deduced amino acid sequence of CbSqS predicted the presence of conserved isoprenoid family domain and catalytic sites. Phylogenetic analysis revealed that CbSqS is closer to Glycine max and monocotyledonous plants. 3D structure prediction using various programs showed CbSqS structure to be similar to SqS from other species. C-terminus truncated recombinant squalene synthase (TruncCbSqS) was expressed in E. coli M15 cells with optimum expression induced with 1 mM IPTG at 37 °C. The gene expression level was analyzed through semi-quantitative RT-PCR and was found to be higher in leaves as compared to the roots.
authors: Department of Biotechnology, Panjab University, Chandigarh 160014, India. shikhakalra11@gmail.com
authors: Shikha
authors: S
authors: Kalra
authors: Sunil
authors: S
authors: Kumar
authors: Neha
authors: N
authors: Lakhanpal
authors: Jagdeep
authors: J
authors: Kaur
authors: Kashmir
authors: K
authors: Singh
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D013186:Farnesyl-Diphosphate Farnesyltransferase / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D019668:Liliaceae / Q000201:enzymology* / Q000235:genetics*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010802:Phylogeny; D018515:Plant Leaves / Q000737:chemistry / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D018517:Plant Roots / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D010940:Plant Proteins; D011994:Recombinant Proteins; D013186:Farnesyl-Diphosphate Farnesyltransferase