Cloning and sequence analysis of L-lysine decarboxylase gene in Huperzia serrata var. longipetiolata / 中草药

ABSTRACT

Objective: To obtain and analyze the full-length L-lysine decarboxylase (LDC) gene sequence of Huperzia serrata var. longipetiolata and predictively analyze its protein structure on the basis of cloning the coding region of LDC gene from four species of Huperziaceae. The species are Huperzia serrata var. longipetiolata, Phlegariurus minchegensis, Phlegariurus austrosinicus, and Phlegariurus petiolatus. Methods: The LDC coding region sequences were cloned by RT-PCR strategy with the template of total RNA extracted from the leaves. Then the sequences were analyzed by means of BLAST and MEGA 5.0. The full-length of LDC gene sequence of H. serrata var. longipetiolata was obtained by RACE technology. And then the secondary structure and three-dimensional structure of LDC protein were predictively analyzed. Results: The coding region sequences were highly similar to the lysine decarboxylase in the database. And the encoding protein of H. serrata var. longipetiolata was highly similar to the amino acid sequences of H. serrata in NCBI, and with high homology to Selaginella moellendorffii. The full-length LDC gene sequence of H. serrata var. longipetiolata contained 1 266 bp open reading frame and encodes a predicted protein of 403 amino acids. The GenBank accession number for this gene is KF040056. Conclusion: The LDC genes of the four species of Huperziaceae are cloned in this study. The full-length LDC gene sequence of H. serrata var. longipetiolata is obtained and analyzed, and its protein structure is predictively analyzed. The result will provide a foundation for exploring the mechanism of huperzine A biosynthesis in the plants of Huperziaceae.