| In this paper, Phenylalanine ammonia-lyase(PAL) gene was firstly cloned from the young leaves of Perilla by rapid amplification of cDNA ends(RACE) technique and it was named Per PAL. Biology information and expression pattern of the novel gene was studied.The primers of PerPAL were designed based on sequences of other PAL genes, such as Salvia and Agastache. The full-length of PerPAL cDNA was2458bp, containing2136bp open reading frame which encoded a pepitide of712amanio acides, and two non-coding regions,966bp5’UTR and226bp3’UTR. The result of BLAST comparision revealed that PerPAL sequence shared90%,85%and82%indentity with Salvia, Agastache and Astragalus, respectively. And the amino acid sequence shared88.69%identity with Salvia ande Agastache. This revealed the PerPAL was cloned successfully.The PerPAL sequence was analysised by bioinformatics software. The result showed that the amino acids was hydrophobic; the research of signal peptide revealed that there wasn’t signal peptide Restriction Enzyme cutting site, so the signal peptide didn’t exist. Meanwhile, there wasn’t transmembrane domain, either; The secondary structure prediction of PerPAL showed that it contained45.57%alpha helix,11.11%outspread chains and43.32%random curly. The three-dimensional model was created for the further study of the senior structure.Phylogenetic tree analysis revealed that each PAL showed different characteristics obviously, denpending on the genera it belonged to. PerPAL had closer relationship with PALs from Lamiaceae plants than from other plants, and PAL gene was ancient existing early in angiosperms and gymnosperms.Relative expressed quantity of PerPAL-1in roots, stems and leaves was analysised by RT-PCR, β-Actin used as internal standard. The results showed that PerPAL gene was expressed the strongest in leaves and the weakest in stems. It was inferred that PALs and the synthesis of rosmarinic acid may be positively correlated in the tissues of Perilla. |
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