Cloning and Sequence Analysis of POD Gene from Pomegranate Peel

Feng Lijuan; Jiao Qiqing; Yin Yanlei; Yang Xuemei; Cheng Yun

doi:10.14083/j.issn.1001-4942.2017.03.003

Abstract：The cDNA sequence of POD was cloned from Taishanhong pomegranate peel by RT-PCR and RACE technique.The nucleotide sequence of POD was analyzed by BLAST and ORF Finder.The amino acid sequence of POD was analyzed by ProtParam,SPOMA and DANMAN softwares.The results indicated that 1 092 bp cDNA sequence of POD was obtained.It contained 828 bp CDS sequence and encoded 276 amino acids.The similarity of POD nucleotide sequence in pomegranate with Nicotiana tabacum,Rubia cordifolia,Arachis ipaensis and Solanum pennellii was 77％,77％,76％ and 76％,respectively.Based on the deduced amino acid sequences of POD genes,the similarity of pomegranate with Litchi chinensis,Citrus clementina,Malus domestica and Pyrus bretschneideri was 89％,87％,84％ and 84％,respectively.The GenBank accession number was KY129694.The theoretical molecular mass of POD protein was 30 582.59 D,and the isoelectric point was 8.83.The protein of POD gene was hydrophilic protein,which had the complete conservative structure domain of peroxidase,and contained two conservative functional domains,Cd00693 and Cl00196.The secondary structure of POD protein was composed by random coil (38.41％),α-helix (37.68％),extended strand (13.77％) and β-turn (10.14％).The results established a theoretical foundation for revealing the mechanism of peel browning in pomegranate fruit.