Cloning and bioinformatic analysis and expression analysis of beta-glucuronidase in Scutellaria baicalensis.
- Author:
Shuang-shuang GUO
;
Lin CHENG
;
Li-min YANG
;
Mei HAN
- Publication Type:Journal Article
- MeSH:
Amino Acid Sequence;
Base Sequence;
Cloning, Molecular;
Computational Biology;
Glucuronidase;
chemistry;
genetics;
metabolism;
Molecular Sequence Data;
Open Reading Frames;
Phylogeny;
Plant Proteins;
chemistry;
genetics;
metabolism;
Protein Structure, Secondary;
Scutellaria baicalensis;
chemistry;
enzymology;
genetics;
Sequence Alignment
- From:
China Journal of Chinese Materia Medica
2015;40(22):4370-4377
- CountryChina
- Language:Chinese
-
Abstract:
The β-Glucuronidase gene (sbGUS) cDNA firstly from Scutellari abaicalensis leaf was cloned by RT-PCR, with GenBank accession number KR364726. The full length cDNA of sbGUS was 1 584 bp with an open reading frame (ORF), encoding an unstable protein with 527 amino acids. The bioinformatic analysis showed that the sbGUS encoding protein had isoelectric point (pI) of 5.55 and a calculated molecular weight about 58.724 8 kDa, with a transmembrane regions and signal peptide, had conserved domains of glycoside hydrolase super family and unintegrated trans-glycosidase catalytic structure. In the secondary structure, the percentage of alpha helix, extended strand, β-extended and random coil were 25.62%, 28.84%, 13.28% and 32.26%, respectively. The homologous analysis indicated the nucleotide sequence 98.93% similarity and the amino acid sequence 98.29% similarity with S. baicalensis (BAA97804.1), in the nine positions were different. The expression level of sGUS was the highest in root based on a real-time PCR analysis, followed by flower and stem, and the lowest was in stem. The results provide a foundation for exploring the molecular function of sbGUS involved in baicalcin biosynthesis based on synthetic biology approach in S. baicalensis plants.
