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Bulletin of Botanical Research ›› 2014, Vol. 34 ›› Issue (6): 816-823.doi: 10.7525/j.issn.1673-5102.2014.06.017

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Cloning and Bioinformation Analysis of PLA Gene Family in Lycium barbarum L.

    

  1. 1.Key Laboratory of Forest Plant Ecology,Ministry of Education,Northeast Forestry University,Harbin 150040;
    2.Ningxia Centers for Diseases Prevention Control,Yinchuan 750004;
    3.College of Biological Science and Engineering,Beifang University of Nationalities,Yinchuan 750021;
    4.School of Laboratory Medicine and Life Science,Wenzhou Medical College,Wenzhou 325035
  • Received:2014-03-15 Online:2014-11-20 Published:2015-01-06
  • Supported by:
     

Abstract: Phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) is a key regulatory enzyme that link primary and secondary metabolism in plants by catalyzing the conversion of L-phenylalanine to cinnamic acid. Four partial sequences of PAL in Lycium barbarum L. were successfully cloned by RT-PCR using a sequence homology strategy, and the bioinformation analysis were carried out. The cDNA fragment of PAL gene family was 1 662-1 683 bp in length, which encodes proteins of 554-561 amino acids with the predicted molecular mass of 60.681-61.250 kD. The amino acid sequence variation between the PAL gene families is 0.005%-0.336%. The estimated isoelectric point (pI) of the putative protein is from 7.02 to 7.71. According to the amino acid sequence and structural analysis, it showed that this protein family contained one conserved domain, namely the shielding domain, and one active site which is phenylalanine/histidine ammonia-lyases. Subcellular localizations of LyPAL1-3 proteins were likely in the mitochondrion and LyPAL4 in the cytoplasm. The secondary structures and tertiary structures of LyPAL1 were abundant in alpha helixs (306) and random coils (179), while were less in beta turns (30) and extended strains (46). Phylogenetic analysis showed that the genes of LyPAL1-3 in L.barbarum L. were closely related to PAL in Capsicum annuum, while LyPAL4 was closely related to PAL in Petunia Juss. (Petunia axillaris, Petunia x hybrid, P.exserta). These four sequences have been registered in GenBank with the accession numbers KC75915-KC759156. Four LyPAL were cloned by molecular biology technology, which would contribute to the molecular mechanisms of the formation of flavonoids and other secondary metabolites in L.barbarum L..

Key words: Lycium barbarum L., PLA gene, cloning, bioinformation analysis

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