LIN Mao-yi, ZHENG Liu, LIU Jin-jie, et al. Cloning and Bioinformatics Analysis of Phenylalanine Ammonia-lyase Gene in [J]. Chinese journal of experimental traditional medical formulae, 2018, 24(1): 38-43.
DOI:
LIN Mao-yi, ZHENG Liu, LIU Jin-jie, et al. Cloning and Bioinformatics Analysis of Phenylalanine Ammonia-lyase Gene in [J]. Chinese journal of experimental traditional medical formulae, 2018, 24(1): 38-43. DOI: 10.13422/j.cnki.syfjx.2018010038.
Cloning and Bioinformatics Analysis of Phenylalanine Ammonia-lyase Gene in
Objective:This study is aimed to clone the key enzyme phenylalanine ammonia-lyase (PAL) from Asarum sieboldii and analyze the sequence characteristics by bioinformatics tools. Method:AsPAL gene was cloned by RT-PCR and RACE strategy by using homologous cloning principle
with the RNA extracted from leaves as the template. The bioinformatics of this gene and its corresponding protein was then analyzed by DNAMAN software and ExPASy online analysis tool. Result:The ORF of AsPAL consisted of 2 157 bp (GenBank accession number:KY428932)
encoding 718 amino acid polypeptides. The molecular weight and theoretical isoelectric point of the deduced AsPAL protein were 78 758 Da and 6.24 respectively. Bioinformatics prediction and analysis indicated that AsPAL protein contained the active center sequence GTITASGDLVPLSYIAG
without transmembrane region and signal peptides. Multiple comparisons of amino acid sequences showed that AsPAL had more than 80% homology with PAL of Cinnamomum osmophloeum
Vitis vinifera
Angelica sinensis and Prunus armeniaca. Conclusion:This study cloned and analyzed PAL from A. sieboldii. The preliminary bioinformatic analysis might be used to establish an experimental basis for exploring PAL gene function and further regulation of methyleugenol biosynthesis from A. sieboladii.