Cloning and bioinformation analysis of chalcone synthase gene from Lycium barbarum in Ningxia / 中草药

ABSTRACT

Objective: To obtain cDNA of chalcone synthase (CHS, EC 2.3.1.74) involved in the flavonoid/isoflavonoid biosynthesis pathway. Methods: The partial sequence of CHS in Lycium barbarum (LyCHS) was successfully cloned by RT-PCR and using a sequence homology strategy, and the bioinformation analysis was carried out. Results: The cDNA fragment of CHS gene was 1148 bp in length, which encoded a protein of 382 amino acids with the predicted relative molecular weight of 4.168 × 104. The estimated isoelectric point (pI) of the putative protein is 6.22. According to the amino acid sequence and structural analysis, it showed that this protein contained one conserved active site, namely chalcone and stilbene synthases active site. Subcellular localizations of LyCHS proteins were likely in the cytoplasm. The secondary and tertiary structures of LyCHS were abundant in α-helixs (166) and random coils (126), while were less in β-turns (25) and extended strains (65). Phylogenetic analysis showed that the genes of LyCHS were closely related to CHS in Solanum pinnatisectum, S. tuberosum, and Lycopersicon esculentum. The sequences had been registered in GenBank with the accession numbers JQ964237. Conclusion: The present results provide the foundation for the next study of CHS gene about its expression and function in L. barbarum.