Cloning of full-length cDNA of gene encoding dihydrodipicolinate synthase from Carthamus tinctorius and construction of plant expression vector / 中草药

ABSTRACT

Objective: To clone the full-length cDNA sequence of DHDPS gene encoding the key enzyme DHDPS in lysing biosynthesis pathway of Carthamus tinctorius, and to construct the plant expression vector. Methods: The dihydrodipicolinate synthase (DHDPS) gene fragment was acquired according to the sequence of transcriptome in C. tinctorius, and the full-length cDNA sequence of CtDHDPS gene from C. tinctorius seeds was cloned by RT-PCR and RACE technologies. The pBasta-DHDPS plant expression vector was constructed using traditional molecular cloning and recombination technique. Results: Bioinformatics analysis showed that the full-length cDNA of CtDHDPS was 1309 bp, open reading frame was 954 bp, encoding a polypeptide of 317 amino acids, the theoretical isoelectric point of the coded protein was 5.93, and the molecular weight was about 34 750.79. The plant expression vector pBasta-DHDPS was successfully constructed by traditional molecular cloning and recombinant technique. Conclusion: The full-length sequence of CtDHDPS gene is obtained and the plant expression vector is successfully constructed, which lays a foundation for the further study on the mechanism of CtDHDPS in regulation of essential amino acid metabolism.