Full-length CDNA Cloning Of Phospholipase D Gene From Agropyron Mongolicum Keng And Its Bioinformatics Analysis

Full-length cDNA sequence of phospholipase D gene from Agropyron mongolicum keng was acquired by RT-PCR and RACE. The structure of sequence, composition and character of amino acid were analyzed by bioinformatics software on line, and molecular evolution trees of PLD was constructed by MEGA3.1. The results were that, full-length cDNA sequence was composed of 2966bp, including a completed ORF which was composed of 2439bp, ATG was start codon and TAG was termination codon, 5′noncoding region which was composed of 127bp lied in upstream of initiation codon, 3′noncoding region which was composed of 368bp lied in downstream of termination codon; it included a signal added poly(A) and a tail which was composed of 32 adenosine monophosphate; The PLD shared 80%～89% amino acid sequence homology to lolium temulentum, zea mays and oryza sativa; its relative molecular weight was 92079.2Da, isoelectric point was 5.24, was a kind of hydrophilic non-transmembrane protein, and mainly located in cytoplasm; The secondary structure of PLD was mainly composed of andom coil; tertiary structure of PLD was formed by C2 domin which next to N terminal and two HKD motifs which was notate sequence of PLD; the molecular evolution trees show that PLD of A. mongolicum keng had higher genetic relationship with lolium temulentum than zea mays and oryza sativa. In this study, we cloned full-length cDNA of PLD gene from A. mongolicum keng and analysed by bioinformatics, the results provide support for studing structure, expression and regulatory mechanism of PLD, and provide theoretical support for screening resistance breeding and controlling land desertification and improving the genetic background of plant.